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US20030068325A1 - Immunogenic peptide composition for the prevention and treatment of Altzheimers Disease - Google Patents

Immunogenic peptide composition for the prevention and treatment of Altzheimers Disease Download PDF

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US20030068325A1
US20030068325A1 US09/865,294 US86529401A US2003068325A1 US 20030068325 A1 US20030068325 A1 US 20030068325A1 US 86529401 A US86529401 A US 86529401A US 2003068325 A1 US2003068325 A1 US 2003068325A1
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peptide
composition
immunogen
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US6906169B2 (en
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Chang Wang
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United Neuroscience Ltd Ireland
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Priority to TW091105087A priority patent/TWI252233B/en
Priority to CN028106210A priority patent/CN1568329B/en
Priority to BR0210010-0 priority patent/BRPI0210010B8/en
Priority to AT09168110T priority patent/ATE496937T1/en
Priority to EP02731223A priority patent/EP1497313B1/en
Priority to ES02731223T priority patent/ES2348975T3/en
Priority to JP2002592863A priority patent/JP4440544B2/en
Priority to CA002448171A priority patent/CA2448171A1/en
Priority to DE60237044T priority patent/DE60237044D1/en
Priority to AT02731223T priority patent/ATE474000T1/en
Priority to DK09168110.6T priority patent/DK2123671T3/en
Priority to MXPA03010631A priority patent/MXPA03010631A/en
Priority to DK02731223.0T priority patent/DK1497313T3/en
Priority to CN2008101692748A priority patent/CN101372511B/en
Priority to EP09168110A priority patent/EP2123671B1/en
Priority to ES09168110T priority patent/ES2360465T3/en
Priority to DE60239093T priority patent/DE60239093D1/en
Priority to AU2002303211A priority patent/AU2002303211B2/en
Priority to CA2665748A priority patent/CA2665748C/en
Priority to PCT/US2002/010293 priority patent/WO2002096350A2/en
Publication of US20030068325A1 publication Critical patent/US20030068325A1/en
Priority to ZA2003/08767A priority patent/ZA200308767B/en
Priority to US10/861,614 priority patent/US7951909B2/en
Assigned to UNITED BIOMEDICAL, INC. reassignment UNITED BIOMEDICAL, INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: WANG, CHANG YI
Priority to HK05102035.8A priority patent/HK1068215A1/en
Priority to HK09107043.3A priority patent/HK1127898A1/en
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Priority to JP2009123671A priority patent/JP5300593B2/en
Priority to JP2009123670A priority patent/JP5300592B2/en
Priority to US12/960,125 priority patent/US8232373B2/en
Assigned to United Neuroscience Limited reassignment United Neuroscience Limited ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: UNITED BIOMEDICAL, INC.
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4711—Alzheimer's disease; Amyloid plaque core protein
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
    • Y10S530/806—Antigenic peptides or proteins
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
    • Y10S530/81—Carrier - bound or immobilized peptides or proteins and the preparation thereof, e.g. biological cell or cell fragment as carrier

Definitions

  • the present invention relates to a composition comprsing a peptide immunogen useful for the prevention and treatment of Alzheimer's Disease.
  • the peptide immunogen comprises a main functional/regulatory site, an N-terminal fragment of Amyloid ⁇ (A ⁇ ) peptide linked to a helper T cell epitope (Th) having multiple class II MHC binding motifs.
  • the peptide immunogen elicit a site-directed immune response against the main functional/regulatory site of the A ⁇ peptide and generate antibodies, which are highly cross-reactive to the soluble A ⁇ 1-42 peptide and the amyloid plaques formed in the brain of Alzheimer's Disease patients.
  • composition of the invention comprising the peptide immunogen is useful for the prevention and treatment of Alzheimer's Disease.
  • AD Alzheimer's Disease
  • ARICEPT® an acetylcholinesterase inhibitor has been approved by the FDA for decelerating the rate of decline of Alzheimer patients. However, it is effective only for a limited period of time and in some patients. Up to the present there is no definitive treatment or cure for this devastating disease.
  • NFT neurofibrillary tangles
  • senile amyloid plaques Two microscopic deposits, i.e., neurofibrillary tangles (NFT) and senile amyloid plaques, were identified by Alois Alzheimer as the pathologic hallmarks of the disease.
  • the neurofibrillary tangles consist of two 10 nm wide filaments twisted around each other, referred to as paired helical filaments (PHFs), a major component of which is phosphorylated tau.
  • PHFs paired helical filaments
  • the phosphorylation of serine at amino acid 262 of the tau protein represents a crucial step leading to physiological dysfunction of tau.
  • PHFs are intracellular and are found in many of the abnormal dendritic and axonal processes, or neurites that make up the periphery of senile amyloid plaques.
  • the senile amyloid plaques consist of disorganized neurophil filaments in an area of up to 150 ⁇ m in cross section with an extra-cellular core of amyloid deposit.
  • the cerebral amyloid plaques are ultrastructurally distinct from PHFs and consist of 4-8 nm wide filaments that are not wound together in pairs.
  • the plaque core consists of aggregates of a peptide, initially referred to as A4, with a relative molecular mass (M) of about 4,000 (Masters et a1 ., Proc Natl Acad Sci USA, 1985, 82:4245-4249).
  • a ⁇ 1-42 has been hypothesized to be related to AD for a number of reasons. Firstly, in peripheral amyloidoses, e.g., primary light chain disease or secondary AA amyloidosis, large amyloid burdens strongly correlate with tissue and organ dysfunction. Secondly, amyloid plaque density positively correlates with premortem dementia scores in AD. Thirdly, A ⁇ 1-42 deposition is the earliest neuropathological marker in AD and related disorders such as Down's syndrome, where it can precede NFT formation by 2-3 decades. Fourthly, 13-amyloidosis is relatively specific to AD and related disorders. Fifthly, A ⁇ 1-42 is toxic to neurons (Yankner et al., Science, 1990; 250:279-282).
  • missense mutations in the structural amyloid precursor protein (APP) gene cause early onset of familial AD (Goate et al., Nature, 1991; 349:704-706; Mullan et al., Nature Genetics, 1992; 1:345-347). Notably, one such mutation causes dramatic A ⁇ 1-42 overproduction (Citron et al., Nature, 1992; 360:672-674).
  • APP structural amyloid precursor protein
  • Kang et al. Nature, 1987; 325:733-737
  • three other groups see 1987 status reports by Anderton, Nature, 1987; 325:658-659 and Barnes, Science, 1987; 235:846-847) independently cloned the gene from which A ⁇ 1-42 is derived.
  • This gene now known as the amyloid precursor protein (APP), encodes a protein of 695 amino-acid residues with a MW of about 79,000 that is expressed in virtually all tissues.
  • APP amyloid precursor protein
  • AD Alzheimer's disease
  • the adjuvant used was for the first injection was Complete Freund's Adjuvant with the incomplete Freund's Adjuvant for the second injection.
  • the adjuvants used are entirely unsuitable for use in humans.
  • the levels of antibodies generated were too low to be effective despite the use of these harsh adjuvants.
  • AD patients amyloid deposits often form at a distance from the site of neuron damage.
  • the best correlation with pathological dementia is loss of synaptic terminals.
  • the loss of synaptic terminals correlates poorly with amyloid load. If the manifestations of disease correlate weakly with amyloid load, then what is the role of A ⁇ ?
  • the article by Klein et al, titled “Targeting small A ⁇ 1-42 oligomers: the solution to an Alzheimer's disease conumdrum?” ( Trends in Neurosciences , 2001; 24:219-224) suggests that fibrils are not the only toxic form of A ⁇ , and perhaps not the neurotoxin that is most relevant to AD.
  • Small oligomers and protofibrils also termed as A ⁇ 1-42 derived diffusible ligands (ADDLs) may also have potent toxic neurological activity.
  • ADDLs A ⁇ 1-42 derived diffusible ligands
  • An AD vaccine for successful immunological intervention will require an immunogen designed to elicit site-directed high affinity antibodies that bind to the senile plaques in the brain tissue to accelerate the clearance of the plaque by the Glial cells, and immunoneutralize the soluble A ⁇ -derived toxins.
  • WO99/27944 disclosed A ⁇ 1-42 or active fragments of A ⁇ 1-42 conjugated to a carrier molecule such as cholera toxin as the active vaccine component. See page 4 of WO 99/27944.
  • page 5 taught that a pharmaceutical composition comprising the immunogen should be free of Complete Freund's Adjuvant [CFA], the only examples showing the efficacy of the A ⁇ 1-42 vaccine for the treatment of AD in transgenic mice employed large doses of aggregated A ⁇ 1-42 peptide in CFA.
  • CFA Complete Freund's Adjuvant
  • Example IX the immune responses in mice to different adjuvants were studied.
  • the adjuvants: MPL, Alum, QS21, and CFA/ICFA were used with the purportedly potent immunogen AN1792 (i.e., aggregated human A ⁇ 1-42 )
  • the level of antibodies to A ⁇ 1-4 were reduced at a statistically significant level in comparison to mice that received the CFA/ICFA vaccines. See, Table 9, and pages 59-64 of WO 99/27944.
  • a ⁇ 1-42 peptide fragments were used (human A ⁇ 1-42 peptides of amino acids 1-5,1-12, 13-28, and 33-42), each fragment was conjugated to sheep anti-mouse IgG as the protein carrier.
  • sheep anti-mouse IgG the protein carrier.
  • the efficacy of antibodies to A ⁇ peptide fragments could only be shown by passive immunization with monoclonal antibodies (Bard et al., Nature Medicine 2000; 6:916-919).
  • the efficacy of these fragments conjugated to sheep anti-mouse IgG was not shown. Therefore, the only immunogen shown to be effective was the aggregated A ⁇ 1-42 peptide in CFA/ICFA.
  • a ⁇ 1-42 peptide or its fragments are self molecules when administered to humans. Therefore, they are less immunogenic or non-immunogenic in humans. It is, thus, necessary to develop clinically acceptable vaccine formulations for administration in humans.
  • promiscuous Th epitopes may be employed to evoke efficient T cell help and may be combined with poorly immunogenic B cell epitopes to provide potent immunogens.
  • Well-designed promiscuous Th/B cell epitopes chimeric peptides have been shown to be useful in eliciting Th responses and resultant antibody responses in most members of a genetically diverse population expressing diverse MHC haplotypes.
  • Promiscuous Th from a number of pathogens, such as measles virus F protein and hepatitis B virus surface antigen, are known.
  • Tables 1 and 2 lists many of the known promiscuous Th that have been shown to be effective in potentiating a short poorly immunogenic peptide, the decapeptide hormone LHRH (U.S. Pat. No. 5,759,551, and 6,025,468).
  • Th epitopes range in size from approximately 15-40 amino acid residues in length, often share common structural features, and may contain specific landmark sequences.
  • a common feature of a Th is that it contains amphipathic helices, alpha-helical structures with hydrophobic amino acid residues dominating one face of the helix and with charged and polar residues dominating the surrounding faces (Cease et al., Proc Natl Acad Sci USA, 1987; 84: 4249-4253).
  • Th epitopes frequently contain additional primary amino acid patterns such as a Gly or charged residue followed by two to three hydrophobic residues, followed in turn by a charged or polar residue. This pattern defines what are called Rothbard sequences.
  • Th epitopes often obey the 1, 4, 5, 8 rule, where a positively charged residue is followed by hydrophobic residues at the fourth, fifth and eighth positions after the charged residue. Since all of these structures are composed of common hydrophobic, charged and polar amino acids, each structure can exist simultaneously within a single Th epitope (Partidos et al., J Gen Virol, 1991; 72:1293). Most, if not all, of the promiscuous T cell epitopes fit at least one of the periodicities described above. These features may be incorporated into the designs of idealized artificial Th sites, including combinatorial Th epitopes.
  • the anitbodies generated by binding to the A ⁇ 1-42 peptide and the senile plaques is expected to accelerate the clearance of these plaques from the brain, promote fibril disaggregation, inhibit fibrillar aggregation, and immunoneutralization of the “soluble A ⁇ -derived toxins” [also termed as A ⁇ -derived diffusible ligands or ADDLs].
  • FIGS. 1 a , and 1 d show significant binding of antibodies to both senile plaques and A ⁇ plaques (both labelled as “P”) on thioflavine S positive blood vessels (labelled as “BV”).
  • FIGS. 1 b and 1 e show the cross reactivity of antibodies raised against the same peptide immunogen in CFA/ICFA.
  • FIGS. 1 c and 1 f show brain sections using preimmune sera.
  • FIGS. 2 a , 2 b , 2 c , 2 d , and 2 e are photographs showing Immunoperoxidase staining of serial sections of AD brain with immune and preimmune sera at 1:100 dilution and under 40 ⁇ magnification.
  • FIGS. 2 a and 2 d showed that the antibodies in guinea pigs immunized with A ⁇ 1-28 - ⁇ K-MVF Th-1-16 (SEQ ID NO:74) prepared in ISA51 water-in-oil emulsion strongly stained the plaques (P) forming a pattern of cores.
  • FIG. 2 b is a photograph of the staining pattern of AD pig brain sections using the same immunogen in CFA/ICFA formulation.
  • FIG. 2 c is a photograph of a guinea pig brain section with preimmune serum and showed no staining.
  • FIG. 2 e shows the brain section with hyperimmune sera generated by immunization with A ⁇ 1-28 peptide alone in CFA/ICFA showing a surprisingly weak staining pattern despite the strong reactivity with A ⁇ 1-28 by ELISA.
  • the present invention relates to an immunogenic composition
  • an immunogenic composition comprising synthetic peptides capable of inducing antibodies against the main functional/regulatory site of the A ⁇ peptide with high cross-reactivity both to the soluble A ⁇ 1-42 peptide and the plaques in the brain of Alzheimer's Disease (AD) patients.
  • the immunogenic composition when administered to an AD patient or a person predisposed to AD is expected to accelerate the clearance of amyloid plaques and immunoneutralization of the soluble A ⁇ derived toxins in the brain to prevent and treat AD.
  • a peptide immunogen of this invention comprise a Th epitope selected from the group consisting of SEQ ID NOS: 1-64 and the immunologically functional analogs thereof linked to a short N-terminal A ⁇ 1-42 peptide fragment selected from the group consisting of 10 to 28 amino acid residues comprising EFRH of the A ⁇ 1-42 peptide, SEQ ID NO:65, or an immunologically functional analog of the A ⁇ 1-42 peptide fragment.
  • the A ⁇ 1-42 peptide fragment is selected from the group SEQ ID NOS: 66-69 or a immunologically functional analogs thereof.
  • the present invention further provides an immunogenic composition
  • an immunologically effective amount of a peptide composition in a pharmaceutically acceptable vaccine formulation comprising an adjuvant or emulsifier selected from the group consisting of liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montanide ISA51, ISA35, ISA206 and ISA 720 as well as the other efficacious adjuvants and emulsifiers.
  • an adjuvant or emulsifier selected from the group consisting of liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montanide ISA51, ISA35, ISA206 and ISA 720 as well as the other efficacious adjuvants and emulsifiers.
  • the present invention further provides a method for the induction of accelerated clearance of amyloid plaques and immunoneutralization of the soluble A ⁇ -derived toxins in the brain to prevent and treat Alzheimer Disease in a mammal by administering one or more of the immunogenic peptides to the mammal for a time and under conditions sufficient to induce antibodies directed against the functional/regulatory site of the A ⁇ 1-42 peptide.
  • a typical example of a vaccine of the present invention is a peptide composition comprising 5-1000 ⁇ g of the peptide immunogen in a vaccine formulated as a water in oil emulsion in a pharmaceutically acceptable adjuvant and/or carrier.
  • a typical method of administering the vaccine is to inject intramuscularly the vaccine formulation at 0.5-2 mL per dose on an immunization schedule of 0, 4, and 8 weeks intervals.
  • Yet another aspect of the invention relates to an immunogenic synthetic peptide of about 30 to about 60 amino acids consisting of a helper T cell (Th) epitope, linked to an N-terminal A ⁇ 1-42 peptide fragment selected from the group consisting of 10 to 28 amino acids with each fragment comprising amino acid residue 1 of the A ⁇ 1-42 peptide.
  • a helper T cell (Th) epitope linked to an N-terminal A ⁇ 1-42 peptide fragment selected from the group consisting of 10 to 28 amino acids with each fragment comprising amino acid residue 1 of the A ⁇ 1-42 peptide.
  • D helper T cell
  • amino acid residue 1 amino acid residue 1
  • the N terminal A ⁇ 1-42 peptide fragment is selected from the group SEQ ID NOs: 66-69 or a peptide analog of N-terminal fragment of A ⁇ 1-42 peptide.
  • amino acid spacers to separate the immunogenic domains may be included.
  • the immunogenic domain elements separated by spacers can be covalently joined in any order provided that either the immunoreactivity of the peptide hapten is substantially preserved or that immunoreactivity to the N-terminal A ⁇ peptide fragment, soluble A ⁇ 1-42 peptide, and the plaques is generated.
  • Th T helper cell epitopes
  • Th T helper cell epitopes
  • Such Th is most reliably supplied to the peptide immunogen by foreign Th epitopes placed on a separate Th peptide domain element that are extrinsic to the target A ⁇ peptide.
  • Such peptide immunogens may be produced as hybrid polypeptides by recombinant DNA expression. They may also be more simply and less expensively supplied as a synthetic peptide immunogen comprising the target hapten B cell site from A ⁇ peptide and T-helper epitopes (Th) appropriate for the host.
  • Such peptides react with helper T-cell receptors and the class II MHC molecules, in addition to antibody binding sites (Babbitt et al., Nature, 1985; 317:359) and thus stimulate a tightly site-specific antibody response to the target antibody binding site.
  • Th was supplied for workable A ⁇ 1-42 peptide immunogens by Th intrinsic to aggregated full length A ⁇ peptide (WO 99/66957; WO 1999/27944; Janus et al., 2000, Morgan et al., 2000) and can be supplied by carrier protein.
  • Th was supplied for workable A ⁇ 1-42 peptide immunogens by Th intrinsic to aggregated full length A ⁇ peptide (WO 99/66957; WO 1999/27944; Janus et al., 2000, Morgan et al., 2000) and can be supplied by carrier protein.
  • a wholly synthetic peptide immunogen enjoys the following advantages over A ⁇ 1-42 peptide aggregates, carrier conjugates and recombinant polypeptides in that the product
  • Immunogenicity of synthetic N-terminal functional-site directed A ⁇ peptide immunogens can be optimized by (1) combining N-terminal A ⁇ 1-42 peptide fragment with selected foreign promiscuous Th sites to which the majority of a population are responsive; and (2) combining A ⁇ peptide fragment with Th whose repertoire is enlarged through combinatorial chemistry, and thereby accommodate to the variable immune responsiveness of a genetically diverse population.
  • peptides composition of the present invention are effective in stimulating the production of antibodies against the main functional/regulatory site of the A ⁇ peptide, with cross-reactivities to the soluble A ⁇ 1-42 and the plaques in the brains of AD patients.
  • the peptide immunogens of the present invention formulated appropriately are effective in humans. It is to be noted that the data obtained in baboons are particularly significant in that this is a species whose immune response closely resemble those of humans.
  • This invention is directed to a novel peptide composition for the generation of high titer polyclonal antibodies with specificity for the main functional/regulatory site of the A ⁇ peptide, with cross-reactivities to the soluble A ⁇ 1-42 and the plaques in the brain of Alzheimer Disease (AD) patients.
  • the antibodies generated by the peptide composition are highly site-specific and bind to the A ⁇ peptides and to amyloids plaques in the brain.
  • the present invention provides an effective method for accelerating the clearance of amyloid plaques and immunoneutralization of soluble A ⁇ derived toxins in the brains for the prevention and treatment of AD.
  • N-terminal A ⁇ 1-42 peptide fragments selected from the group consisting of 10 to 28 amino acids wherein each fragment comprises EFRH of the A ⁇ 1-42 peptide (SEQ ID NO:65), are short linear peptides which, by themselves are non-immunogenic.
  • the short A ⁇ 1-42 peptide fragments can be immuno-potentiated by chemical coupling to a carrier protein, for example, keyhole limpet hemocyanin (KLH) or by fusion to a carrier polypeptide through recombinant DNA expression, for example, hepatitis B surface antigen.
  • KLH keyhole limpet hemocyanin
  • the deficiency of such “A ⁇ peptide(s)-carrier protein” vaccines is that a major portion of antibodies generated are non-functional antibodies directed against the carrier protein.
  • the immunogens of the present invention are wholly synthetic peptide immunogens comprising N-terminal fragment of A ⁇ 1-42 peptide of 10 to 28 amino acids with each fragment comprising EFRH of the A ⁇ 1-42 peptide covalently linked to promiscuous Th epitopes selected from the group consisting of SEQ ID NOs: 1 to 64.
  • the immunogens of the invention elicit the production of site-specific antibodies which bind to the A ⁇ 1-42 peptide and its aggregates and are cross reactive with amyloid plaques in the brain to provide for accelerated clearance of amyloid plaques and immunoneutralization of the soluble A ⁇ -derived toxins in the brain.
  • the immunogen of the present invention is useful in preventing and treating AD.
  • helper T cell epitopes (Th) useful in the invention comprise multiple class II MHC binding motifs. Specific examples of Th covalently linked to an N-terminal A ⁇ 1-42 peptide fragment are provided.
  • the results of anti-sera from animals immunized with the immunogen peptides of the present invention demonstrate that potent site-directed A ⁇ peptide reactive antibodies are generated, in a genetically diverse host population.
  • the synthetic immunogenic peptide of the present invention are approximately 20 to 100 amino acids long and comprise:
  • helper T cell (Th) epitope selected from the group consisting of SEQ ID Nos: 1 to 64;
  • (iii) optionally a spacer consisting of at least an amino acid to separate the immunogenic domains.
  • the N terminal fragment of the A ⁇ 1-42 peptide is selected from the group consisting of SEQ ID NOS: 66-69 and an immunologically effective analog thereof.
  • the Th peptide is covalently attached to either the N— or C-terminus of the target N-terminal fragment of A ⁇ 1-42 peptide optionally with a spacer (e.g., Gly-Gly, ⁇ -N Lys).
  • the peptide immunogen of this invention is represented by one of the following formula:
  • each A is independently an amino acid
  • each B is a linking group selected from the group consisting of an amino acid, gly-gly, ( ⁇ , ⁇ -N)lys, Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:73);
  • Each Th comprise an amino acid sequence that constitutes a helper T cell epitope, or an immune enhancing analog or segment thereof;
  • N-terminal fragment of A ⁇ 1-42 peptide is a synthetic peptide B cell target site antigen and is a fragment of about 10 to about 28 amino acid residues wherein each fragment comprises EFRH of the A ⁇ 1-42 peptide or an immunologically functional analog thereof;
  • X is an ⁇ -COOH or ⁇ -CONH 2 of an amino acid
  • n is from 0 to about 10;
  • m is from 1 to about 4.
  • o is from 0 to about 10.
  • the peptide immunogen of the present invention comprises from about 20 to about 100 amino acid residues, preferably from about 25 to about 60 amino acid residues.
  • the (N-terminal fragment of A ⁇ 1-42 peptide) is selected from the group consisting of SEQ ID Nos: 66-69 and preferably the Th epitope is selected from the group consisting of SEQ ID NOs: 1, 3, 4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54.
  • A is an amino acid
  • it is a non-naturally occurring or naturally occurring amino acid.
  • Non-naturally occurring amino acids include, but are not limited to, ⁇ -N lysine, ⁇ -alanine, ornithine, norleucine, norvaline, hydroxyproline, thyroxine, ⁇ -amino butyric acid, homoserine, citrulline and the like.
  • Naturally-occurring amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine.
  • n may include a spacer, e.g., Gly-Gly, ⁇ -N Lys.
  • B is a spacer and is an amino acid which can be naturally occurring or the non-naturally occurring amino acids as described above.
  • Each B is independently the same or different.
  • the amino acids of B can also provide a spacer, e.g., Gly-Gly, ⁇ -Lys, or lysine between the promiscuous Th epitope and the N-terminal fragment of A ⁇ 1-42 peptide (e.g., SEQ ID NOs:66-69) or an immunologically functional analog thereof.
  • the Gly-Gly or F-Lys spacer can disrupt any artifactual secondary structures created by the joining of the Th epitope with an N-terminal fragment of A ⁇ 1-42 peptide or its immunologically functional analog and thereby eliminate interference between the Th and/or B cell responses.
  • the amino acids of B can also form a spacer which acts as a flexible hinge that enhances separation of the Th and the N-terminal fragments of A ⁇ 1-42 peptide. Examples of sequences encoding flexible hinges are found in the immunoglobulin heavy chain hinge region.
  • Flexible hinge sequences are often proline rich.
  • One particularly useful flexible hinge is provided by the sequence Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:77), where Xaa is any amino acid, and preferably aspartic acid.
  • SEQ ID NO:77 Pro-Pro-Xaa-Pro-Xaa-Pro
  • Xaa is any amino acid, and preferably aspartic acid.
  • the conformational separation provided by the amino acids of B permits more efficient interactions between the presented peptide immunogen and the appropriate Th cells and B cells to enhances the immune responses to the Th epitope and the antibody-eliciting epitope or their immunologically functional analogs.
  • Th is a sequence of amino acids (natural or non-natural amino acids) that comprises a Th epitope.
  • a Th epitope may be a continuous or discontinuous epitope. In a discontinuous Th epitope, not every amino acid of Th is necessary.
  • a Th epitope, or an analog or fragment thereof, is capable of enhancing or stimulating an immune response to the N-terminal fragment of A ⁇ 1-42 peptide.
  • Th epitopes that are immunodominant and promiscuous are highly and broadly reactive across animal and human populations with widely divergent MHC types (Partidos et al., 1991; U.S. Pat. No. 5,759,551).
  • the Th epitope of the subject peptides is about 10 to about 50 amino acids, preferably from about 10 to about 30 amino acids. When multiple Th epitopes are present (i.e., m>2), each Th epitope may be the same or different.
  • a Th segment comprises a contiguous portion of a Th epitope that is sufficient to enhance or stimulate an immune response to the N-terminal fragment of A ⁇ 1-42 peptide.
  • Th epitopes of the present invention include those derived from foreign pathogens including but not limited to those exemplified in Table 1 (SEQ ID Nos:1-21). Further, Th epitopes include idealized artificial Th and artificial idealized combinatorial Th disclosed in WO 99/66957 and listed here in Table 2 as SEQ ID Nos 22-64. Peptides comprising combinatorial Th are produced simultaneously in a single solid-phase peptide synthesis in tandem with the N-terminal fragment of A ⁇ 1-42 peptide, A and B. The Th epitopes also include immunologically functional analogs thereof, having conservative substitutions, additions, deletions and insertions therein of from one to about 10 amino acid residues as long as the Th-stimulating function has not been essentially modified.
  • the Th epitope is covalently attached through a spacer B to either the N terminus or C terminus of the N-terminal fragment of A ⁇ 1-42 peptide or an immunologically functional analog thereof.
  • An immunologically functional analog of the N-terminal fragment of A ⁇ 1-42 peptide may comprise conservative substitutions, additions, deletions, or insertions of from one to about four amino acid residues as long as immune responses that are crossreactive with the A ⁇ 1-42 peptides are elicited.
  • the conservative substitutions, additions, and insertions can be accomplished with natural or non-natural amino acids as defined above.
  • the preferred peptide immunogens of this invention are those comprising the N-terminal fragment of the A ⁇ 1-42 peptide fragments selected from the group consisting of SEQ ID NOs: 66-69 or an immunologically functional analog thereof; a spacer (e.g., Gly-Gly, ⁇ -Lys); a Th epitope selected from the group consisting of an HB s Th (SEQ ID NO:1); HB c Th (SEQ ID NO:20); MVF Th (SEQ ID NOS:8, 9); PT Th (SEQ ID NOs:4, 5, 7), TT Th (SEQ ID NOs:3, 4, 6); CT Th (SEQ ID NOs:12, 21); DT Th (SEQ ID NO:13, 14), MVF Th derived artificial Th selected from the group consisting of SEQ ID Nos:38-40, 47-51); HBV Th derived artificial Th selected from the group consisting of SEQ ID NOS: 52-54. See Tables 1 and 2.
  • Peptide compositions which contain a cocktail of the subject peptide immunogens with two or more Th epitopes may enhance immunoefficacy in a broader population and thus provide an improved immune response to the A ⁇ 1-42 peptides and their fragments.
  • the peptide immunogens of this invention can be made by chemical synthesis methods which are well known to the ordinarily skilled artisan. See, for example, Fields et al., Chapter 3 in Synthetic Peptides: A User's Guide , ed. Grant, W. H. Freeman & Co., New York, N.Y., 1992, p. 77. Hence, peptides can be synthesized using the automated Merrifield techniques of solid phase synthesis with the ⁇ -NH 2 protected by either t-Boc or F-moc chemistry using side chain protected amino acids on, for example, an Applied Biosystems Peptide Synthesizer Model 430A or 431.
  • Preparation of peptide constructs comprising combinatorial library peptides for Th epitopes can be accomplished by providing a mixture of alternative amino acids for coupling at a given variable position. After complete assembly of the desired peptide immunogen, the resin is treated according to standard procedures to cleave the peptide from the resin and deblock the functional groups on the amino acid side chains. The free peptide is purified by HPLC and characterized biochemically, for example, by amino acid analysis or by sequencing. Purification and characterization methods for peptides are well known to one of ordinary skill in the art.
  • the immunogen of the present invention may also be prepared as a branched polymer by synthesis of the desired peptide construct directly onto a branched poly-lysyl core resin (Wang, et al., Science, 1991; 254:285-288).
  • the longer synthetic peptide immunogens can be synthesized by well known recombinant DNA techniques. Such techniques are provided in well-known standard manuals with detailed protocols.
  • To construct a gene encoding a peptide of this invention the amino acid sequence is reverse translated to obtain a nucleic acid sequence encoding the amino acid sequence, preferably with codons that are optimum for the organism in which the gene is to be expressed.
  • a synthetic gene is made, typically by synthesizing oligonucleotides which encode the peptide and any regulatory elements, if necessary.
  • the synthetic gene is inserted in a suitable cloning vector and transfected into a host cell.
  • the peptide is then expressed under suitable conditions appropriate for the selected expression system and host.
  • the peptide is purified and characterized by standard methods.
  • the efficacy of the peptide composition of the present invention can be established by injecting an animal, for example, guinea pigs, with an immunogenic composition comprising peptides of the invention. See, Table 4, SEQ ID NOS:70-75.
  • the humoral immune response to the N-terminal fragment of A ⁇ 1-42 peptide and the soluble A ⁇ 1-42 peptide are monitored. A detailed description of the procedures used is provided in the Examples hereinbelow.
  • Another aspect of this invention provides a peptide composition comprising an immunologically effective amount of one or more of the peptide immunogens of this invention in a pharmaceutically acceptable delivery system.
  • the subject peptide composition can be formulated as a vaccine using pharmaceutically acceptable adjuvants, carriers or other ingredients routinely employed in the formulation of vaccines.
  • ingredients that can be used in this invention are adjuvants or emulsifiers including alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720 as well as the other efficacious adjuvants and emulsifiers.
  • the composition may be formulated for immediate release or sustained release.
  • the composition may also be formulated for induction of systemic immunity, e.g., by entrapment in or coadministration with microparticles. Such formulations are readily available to one of ordinary skill in the art.
  • the immunogens of the present invention can be administered via any conventional route, such as subcutaneous, oral, intramuscular, parenteral or enteral route.
  • the immunogens can be administered in a single dose or in multiple doses.
  • a suitable immunization schedule is readily determined and available to one of ordinary skill in the art.
  • the peptide composition of the present invention comprises an effective amount of one or more of the peptide immunogens of the present invention and a pharmaceutically acceptable carrier.
  • a composition in a suitable dosage unit form generally contains about 0.25 ⁇ g to about 500 ⁇ g of the immunogen per kg body weight.
  • the effective amount may be conveniently divided per dosage unit.
  • an initial dose e.g. 0.0025-0.5 mg per kg body weight; preferably 1-50 ⁇ g per kg of body weight of the peptide immunogen is to be administered by injection, preferably intramuscularly, followed by repeat (booster) doses of a similar amount. Dosage will depend on the age, weight and general health of the subject as is well known in the vaccine and therapeutic arts.
  • the immune response of the synthetic A ⁇ 1-42 peptide immunogens can be improved by delivery through entrapment in or on biodegradable microparticles of the type described by O'Hagan et al. ( Vaccine, 1991; 9: 768-771).
  • the immunogens can be encapsulated with or without an adjuvant in biodegradable microparticles, to potentiate immune responses, and to provide time-controlled release for sustained or periodic responses, and for oral administration, (O'Hagan et al., 1991; and, Eldridge et al., 1991; 28: 287-294).
  • Peptide immunogens listed in Table 4 were synthesized individually by the Merrifield solid-phase synthesis technique on Applied Biosystems automated peptide synthesizers (Models 430, 431 and 433A) using Fmoc chemistry.
  • Preparation of peptide immunogens comprising a combinatorial library Th i.e., idealized artificial Th site such as MvF derived Th1-8 (SEQ ID NOs:38-40), can be accomplished by providing a mixture of the desired amino acids for chemical coupling at a given position as specified in the design.
  • the resin was treated according to standard procedure using trifluoroacetic acid to cleave the peptide from the resin and deblock the protecting groups on the amino acid side chains.
  • the cleaved, extracted and washed peptides were purified by HPLC and characterized by mass spectrometry and reverse phase HPLC.
  • a ⁇ -derived peptide immunogens were evaluated on groups of guinea pigs as specified by the experimental immunization protocol outlined below and by serological assays for determination of immunogenicity.
  • Adjuvants Complete Freund's Adjuvant (CFA)/Incomplete
  • Adjuvant (IFA); or water in oil emulsions unless otherwise specified.
  • CFA/IFA groups received CFA week 0, IFA in subsequent weeks.
  • Assay Specific ELISAs for each immune serum's anti-peptide activity.
  • the Solid phase substrate was the A ⁇ peptide fragment e.g. A ⁇ 1-14 or full length A ⁇ 1-42 (SEQ ID NOs: 67 and 65). Blood was collected and processed into serum, and stored prior to ELISA with the target peptides.
  • the immunoreactivities of the antibodies elicited against A ⁇ peptides and against the soluble A ⁇ 1-42 peptides were determined by ELISAs (enzyme-linked immunosorbent assays) using 96-well flat bottom microtiter plates which were coated with the A ⁇ 1-42 peptide fragments, SEQ ID NOs: 67 or 65 as the immunosorbent. Aliquots (100 ⁇ L) of the peptide immunogen solution at a concentration of 5 ⁇ g/mL were incubated for 1 hour at 37° C. The plates were blocked by another incubation at 37° C. for 1 hour with a 3% gelatin/PBS solution. The blocked plates were then dried and used for the assay.
  • a ⁇ 1-28 was compared to A ⁇ 1-14 , a shorter N-terminal fragment of A ⁇ 1-42 .
  • a more potent adjuvant suitable for human use (Montamide ISA51, Seppic, Paris, FR) was employed for the preparation of a water-in-oil emulsion for formulating the vaccine.
  • the relative immunogenicities for the three A ⁇ peptides i.e. A ⁇ 1-14 , A ⁇ 1-28 and A ⁇ 1-42 ) were ranked A ⁇ 1-28 >A ⁇ 1-42 >A ⁇ 1-14 .
  • the loss of the C-terminal 14 mer from A ⁇ 1-42 improved rather than reduced the immunogenicity.
  • the antibody response against A ⁇ is primarily directed to the N-terminal region, particularly the A ⁇ 1-14 N-terminal fragment as shown by ELISA data (Table 6).
  • a further shortening of the A ⁇ 1-28 fragment from the C-terminal to form the A ⁇ 1-14 fragment resulted in a loss in immunogenicity.
  • the short A ⁇ 1-14 fragment contains the main functional/regulatory site, EFRH, located at positions 3-6 of the A ⁇ 1-42 peptide as reported by Solomon et al.
  • EFRH main functional/regulatory site
  • the blocking of this epitope by antibodies modulates the dynamics of aggregation as well resolubilization of already formed aggregates (Soloman et al., Proc Nat/Acad. Sci, 1996; 93:452-455 ; Proc Natl Aca. Sci, 1997; 94:4109-4112).
  • Most of the anti-A ⁇ 1-28 and A ⁇ 1-42 antibodies are directed against the N-terminal fragment of the A ⁇ 1-42 peptide containing this epitope (Table 6).
  • the A ⁇ 1-14 fragment by itself was poorly immunogenic.
  • the results of this experiment suggest the presence of an intrinsic Th epitope within the A ⁇ 15-28 segment. This intrinsic Th epitope accounts for the modest immunogenicities of A ⁇ 1-28 and A ⁇ 1-42 peptides in
  • Th epitope in the A ⁇ 15-28 fragment is desirable. However, it is desirable to be able to engineer a more potent immunogen for a successful human vaccine when faced with the limitation of a restricted human MHC molecule, the number of appropriate doses and the type of adjuvants permitted for human use. Therefore, we attempted the linkage of a foreign or extrinsic Th such as that derived from HBV Th (SEQ ID NO: 1) to the C-terminal of the A ⁇ 1-28 peptide (SEQ ID NO:66). The extrinsic Th epitope significantly enhanced the immunogenicity of the A ⁇ 1-28 fragment as shown in Table 6.
  • This peptide immunogen represents a peptide immunogen with the formula:
  • A is ⁇ NH 2 , with A ⁇ 1-28 being an N-terminal fragment of A ⁇ 1-42 ;
  • B is glycine
  • Th is a helper T cell epitope derived from a foreign pathogen
  • HBsAg Th (SEQ ID NO: 1), and wherein n is 1, m is 1 and o is 2.
  • A is ⁇ NH 2 , wherein the N-terminal fragment is A ⁇ 1-10 , A ⁇ 1-12 , A ⁇ 1-14 or A ⁇ 1-28 ;
  • B is ⁇ -N Lysine, a spacer linked through its epsilon amino group to the next amino acid;
  • Th is a helper T cell epitope derived from an idealized artificial Th, MVF Th1-16(SEQ ID NO:51), wherein n is 1, m is 1 and o is 1.
  • the useful B cell site derived from A ⁇ 1-42 should be in the size range of about 10 to about 28 residues.
  • the non-immunogenic N-terminal fragment such as A ⁇ 1-14 of A ⁇ peptide was linked either through an ⁇ N-lysine spacer to an artificial Th peptide designated as MVF Th 1-16 (SEQ ID NO:51), or through a standard chemical coupling procedure to a conventional carrier protein KLH.
  • the two immunogenic constructs were evaluated in guinea pigs for their relative “site-directed” immunogenicities to A ⁇ peptide and the resultant respective reactivity of the antibodies towards their respective carriers, the artificial Th epitope or the KLH carrier protein, according to the procedures described in Example 2.
  • the short A ⁇ 1-14 peptide alone as a control immunogen, and the two immunogenic constructs were formulated in a water-in-oil emulsion containing the adjuvant ISA51, a formulation that is suitable for human use.
  • the N-terminal A ⁇ 1-14 fragment by itself is non-immunogenic as expected.
  • the synthetic immunogen comprising A ⁇ 1-14 fragment and artificial Th was found to be highly immunogenic in eliciting site-directed antibodies to A ⁇ 1-14 .
  • the antibodies were also found to be highly cross-reactive to soluble A ⁇ 1-42 peptide as early as 4 weeks after the initial immunization (Log 10 titers of 4.094 and 4.126 for 4 and 6 weeks post initial immunization respectively).
  • these AP-reactive high titer immune sera were tested by ELISA on the MVF Th1-16 peptide (SEQ ID NO 51) coated plate, they were found to be negative (Log 10 titer of 0.038 and 0.064 for 4 and 6 weeks post initial immunization respectively) showing that irrelevant antibodies were not produced.
  • Table 8 clearly demonstrated the highly specific site-directed characteristic of the peptide immunogen of the present invention.
  • the immunogens with the carrier protein KLH was found to be highly -; immunoreactive with the conventional peptide-carrier protein conjugate (e.g. Log 10 .: titers of 4.903 and 5.018 for 4 and 6 weeks post initial immunization respectively).
  • the antibodies elicited were only moderately crossreactive with the soluble A ⁇ 1-42 peptide (e.g. with Log 10 titers of 3.342 and 2.736 for 4 and 6 weeks post initial immunization respectively). This is approximately 1 ⁇ to 100 ⁇ less than SEQ ID NO:73.
  • the peptide immunogens of the present invention were highly site-directed and focused. Only functionally important antibodies towards the anti-aggregation and disaggregation sites on the N-terminal fragment of the A ⁇ peptide were generated rather than towards irrelevant carrier sites.
  • TSBV thioflavine S positive blood vessels
  • FIG. 1 serial cross sections of brains from 2 AD patients were initially examined at 1 ⁇ magnification. Sections (a), (b) and (c) are from AD Brain 1 and (d), (e) and (e are from AD brain 2. Preimmune normal serum and immune sera from guinea pigs collected at 6 weeks post-initial immunization were tested by immunoperoxidase staining on cryostat sections from AD temporal cortex rich in plaques and neurofilament tangles (NFT). The immune sera used in the first study shown on slides FIGS.
  • the antibodies elicited by the vaccine formulated with ISA51 is distinguishable from the antibodies raised by the vaccine formulated in CFA/ICFA.
  • the antibodies generated by the vaccines formulated according to the present invention provided antibodies that have the desired higher cross reactivity to senile plaques in the brain tissue. Preimmune serum gave no staining in corresponding serial sections shown in slides FIGS. 1 c and 1 f.
  • FIGS. 2 a to 2 e Further Immunoperoxidase staining of serial cross sections of AD brain 1 with preimmune and immune sera at 1:100 dilution are shown in FIGS. 2 a to 2 e at 40 ⁇ magnification.
  • the sera obtained from animals immunized with A ⁇ 1-28 - ⁇ K-MVF Th 1-16 (Seq ID NO:74) prepared in ISA 51 water-in-oil emulsion strongly stained the plaques forming a pattern of cores as shown in slides FIGS. 2 a and 2 d .
  • staining with immune sera prepared against the corresponding CFA/ICFA formulation gave a different staining pattern in that reactivities with plaques were predominantly on the blood vessels as shown in FIG.
  • a fourth baboon X798 was given 100 ug/0.5 mL doses of an equimolar mixture of free peptides A ⁇ 1-28 and A ⁇ 1-42 formulated in alum, the standard adjuvant approved for human use. Preimmune sera were used as the negative control.
  • Sera from all four immunized animals were collected and evaluated for their antibody reactivities with the functional site by A 1-14 ELISA, and for reactivities with soluble A ⁇ 1-42 by A ⁇ 1-42 ELISA (for sera collected at 0, 5 and 8 wpi).
  • the cross-reactivities of the anti-sera (8 wpi only) with the senile plaques and the plaques in thioflavine S positive blood vessels were evaluated by immunostaining as described in Example 6.
  • the antibody detector used is an Fab fragment from anti-human IgG that recognizes all human isotypes and is cross-reactive with baboon IgG.
  • a mixture containing two to three synthetic immunogens of the present invention can be used for formulation into vaccines at from about 25 to 1000 ug per dose to elicit functional anti-A ⁇ 1-14 antibodies in genetically diverse human populations for the prevention and treatment of AD.
  • Broad immunogenicity in humans is expected due to the presence of a promiscuous Th epitope in the peptide immunogen of the invention that provides for achieving broad MHC recognition.
  • Th TABLE 1 Pathogen-derived Promiscuous T Helper Cell Epitopes (Th) Description of Th Amino Acid Sequence SEQ ID NO HBs Th a FFLLTRILTIPQSLD 1 PT 1 Th a KKLRRLLYMIYMSGLAVRVHVSKEEQYYDY 2 TT 1 Th a KKQYIKANSKFIGITEL 3 TT 2 Th a KKFNNFTVSFWLRVPKVSASHL 4 PT IA Th a YMSGLAVRVHVSKEE 5 TT 3 Th a YDPNYLRTDSDKDRFLQTMVKLFNRIK 6 PT 2 Th a GAYARCPNGTRALTVAELRGNAEL 7 MVF 1 Th a LSEIKGVIVHRLEGV 8 MVF 2 Th a GILESRGI KARITHVDTESY 9 TT 4 Th a WVRDIIDDFTNESSQKT 10 TT 5 Th a DVSTIVPYIGPALNHV 11 CT Th a ALNI

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Abstract

The present invention relates to a composition comprsing a peptide immunogen useful for the prevention and treatment of Alzheimer's Disease. More particularly, the peptide immunogen comprises a main functional/regulatory site, an N-terminal fragment of Amyloid β (Aβ) peptide linked to a helper T cell epitope (Th) having multiple class II MHC binding motifs. The peptide immunogen elicit a site-directed immune response against the main functional/regulatory site of the Aβ peptide and generate antibodies, which are highly cross-reactive to the soluble Aβ1-42 peptide and the amyloid plaques formed in the brain of Alzheimer's Disease patients. The antibodies elicited being cross reactive to the soluble Aβ1-42 peptide, promote fibril disaggregation and inhibit fibrillar aggregation leading to immunoneutralization of the “soluble Aβ-derived toxins”; and being cross-reactive to the amyloid plaques, accelerate the clearance of these plaques from the brain. Thus, the composition of the invention comprising the peptide immunogen is useful for the prevention and treatment of Alzheimer's Disease.

Description

    FIELD OF THE INVENTION
  • The present invention relates to a composition comprsing a peptide immunogen useful for the prevention and treatment of Alzheimer's Disease. More particularly, the peptide immunogen comprises a main functional/regulatory site, an N-terminal fragment of Amyloid β (Aβ) peptide linked to a helper T cell epitope (Th) having multiple class II MHC binding motifs. The peptide immunogen elicit a site-directed immune response against the main functional/regulatory site of the Aβ peptide and generate antibodies, which are highly cross-reactive to the soluble Aβ[0001] 1-42 peptide and the amyloid plaques formed in the brain of Alzheimer's Disease patients. The antibodies elicited being cross reactive to the soluble Aβ1-42 peptide, promote fibril disaggregation and inhibit fibrillar aggregation leading to immunoneutralization of the “soluble AD-derived toxins”; and being cross-reactive to the amyloid plaques, accelerate the clearance of these plaques from the brain. Thus, the composition of the invention comprising the peptide immunogen is useful for the prevention and treatment of Alzheimer's Disease.
    • BACKGROUND OF THE INVENTION
  • Alzheimer's Disease (AD) is a chronic, neurodegenerative disorder characterized by a loss of cognitive ability and severe behavoral abnormalities in a patient leading to the eventual death of the patient. There are currently 2.5 to 4.0 million AD patients in the U.S. and 17 to 25 million worldwide. It is the fourth leading cause of death in Western cultures, preceded only by heart disease, cancer, and stroke. ARICEPT®, an acetylcholinesterase inhibitor has been approved by the FDA for decelerating the rate of decline of Alzheimer patients. However, it is effective only for a limited period of time and in some patients. Up to the present there is no definitive treatment or cure for this devastating disease. [0002]
  • Two microscopic deposits, i.e., neurofibrillary tangles (NFT) and senile amyloid plaques, were identified by Alois Alzheimer as the pathologic hallmarks of the disease. The neurofibrillary tangles consist of two 10 nm wide filaments twisted around each other, referred to as paired helical filaments (PHFs), a major component of which is phosphorylated tau. The phosphorylation of serine at amino acid 262 of the tau protein represents a crucial step leading to physiological dysfunction of tau. PHFs are intracellular and are found in many of the abnormal dendritic and axonal processes, or neurites that make up the periphery of senile amyloid plaques. The senile amyloid plaques consist of disorganized neurophil filaments in an area of up to 150 μm in cross section with an extra-cellular core of amyloid deposit. The cerebral amyloid plaques are ultrastructurally distinct from PHFs and consist of 4-8 nm wide filaments that are not wound together in pairs. The plaque core consists of aggregates of a peptide, initially referred to as A4, with a relative molecular mass (M) of about 4,000 (Masters et a1[0003] ., Proc Natl Acad Sci USA, 1985, 82:4245-4249).
  • A partial amino acid sequence of A4, now renamed amyloid β peptide (or Aβ[0004] 1-42), shows that it is similar to the amyloid β protein isolated from cerebral blood vessels of patients with Alzheimer's disease or Down's syndrome (Glenner and Wong, Biochem Biophys Res Comm, 1984; 120:885-890; 122:1131-1135).
  • Aβ[0005] 1-42 has been hypothesized to be related to AD for a number of reasons. Firstly, in peripheral amyloidoses, e.g., primary light chain disease or secondary AA amyloidosis, large amyloid burdens strongly correlate with tissue and organ dysfunction. Secondly, amyloid plaque density positively correlates with premortem dementia scores in AD. Thirdly, Aβ1-42 deposition is the earliest neuropathological marker in AD and related disorders such as Down's syndrome, where it can precede NFT formation by 2-3 decades. Fourthly, 13-amyloidosis is relatively specific to AD and related disorders. Fifthly, Aβ1-42 is toxic to neurons (Yankner et al., Science, 1990; 250:279-282). Lastly, missense mutations in the structural amyloid precursor protein (APP) gene cause early onset of familial AD (Goate et al., Nature, 1991; 349:704-706; Mullan et al., Nature Genetics, 1992; 1:345-347). Notably, one such mutation causes dramatic Aβ1-42 overproduction (Citron et al., Nature, 1992; 360:672-674).
  • In 1987, Kang et al. ([0006] Nature, 1987; 325:733-737) and three other groups (see 1987 status reports by Anderton, Nature, 1987; 325:658-659 and Barnes, Science, 1987; 235:846-847) independently cloned the gene from which Aβ1-42 is derived. This gene, now known as the amyloid precursor protein (APP), encodes a protein of 695 amino-acid residues with a MW of about 79,000 that is expressed in virtually all tissues. There are at least five splicing variants of APP, four of which contain the β-amyloid peptide sequence.
  • Four genes have been implicated in familial forms of AD. Three of the genes, βAPP, presenilin 1 and presenilin 2, when mutated, cause autosomal dominant early forms of AD. The fourth gene, Apolipoprotein E, has a naturally occurring polymorphic form, ApoE4, that represents a major genetic risk factor for the development of the disease. The concept that alterations in several distinct genes can lead to a chronic imbalance between Aβ[0007] 1-42 production and its clearance, with the resulting aggregation of first the 42-residue and then the 40-residue peptide into cytotoxic plaques, is supported by available evidence. The evidence strongly suggests that defects in each of these four genes predispose the AD phenotype by (1) enhancing the production and/or the deposition of Aβ1-42 peptides or (2) by decreasing the clearance of ApoE4 from tissue (Selkoe, J Biol Chem, 1996; 271:18295-18298).
  • From available data, it appears that aggregated but not monomeric Aβ[0008] 1-42 peptides can induce cell dysfunction and death in vitro by a range of presumably interrelated mechanisms. These include oxidative injury (Thomas et al., Nature, 1996; 380:168-171; Behl et al., Cell, 1994; 77:817-827), alterations in intracellular calcium homeostasis (Arispe et al., Proc Natl Acad Sci USA, 1993; 90:567-571), and cytoskeletal reorganization (Busciglio et al., Neuron, 1995; 14:879-888). Sufficient knowledge of some of the principal steps in the amyloid-induced cascade has emerged, even though the cascade hypothesis is hotly contested.
  • Pharmalogical approaches of identifying small molecules which could inhibit one or another step of the amyloid induced cascade is now well under way. Of particular interest are two approaches: attempts to interfere with the aggregation of Aβ[0009] 1-42 peptides by decreasing the secretion of Aβ1-42 peptides from neuronal and glial cells or inhibit the toxicity that these extracellular aggregates produce on neurons and glial cells and their processes. A third approach attempts to control the specialized inflammatory response that appears to be triggered by aggregated Aβ1-42 (including microglial stimulation, activation of the classical complement cascade, cytokine release, and reactive astrocytosis) may prove to be of benefit to Alzheimer patients.
  • Aside from the above-mentioned pharmacological approaches for AD intervention, immunological interventions have also been attempted. Soloman et al. ([0010] Proc Natl Acad. Sci, 1996; 93:452-455; Proc Natl Aca. Sci, 1997; 94:4109-4112) showed that three specific monoclonal antibodies, directed toward a site in the N-terminal region of the human Aβ1-42 peptide, bind in varying degrees to preformed fibrils leading to their disaggregation and inhibition of their neurotoxic effect. The antibodies were also found to prevent the formation of fibrillar Aβ1-42. Solomon et al. (WO 01/18169) also attempted to prepare a phage display of an epitope of the Aβ1-42 peptide and administering the phage displayed epitope or peptide containing the epotope intraperitonially to mice to elicit antibodies to the Aβ1-42 peptide. In vitro testing with rat phenochromocytoma showed that a 1:5 dilution of the anitsera prevented the neurotoxicity of Aβ1-42. The antiserum at a dilution of 1:5 and 1:20 was also shown to disrupt the fibril structure of Aβ in vitro with extensive deterioration of fibril morphology. However, the adjuvant used was for the first injection was Complete Freund's Adjuvant with the incomplete Freund's Adjuvant for the second injection. The adjuvants used are entirely unsuitable for use in humans. Moreover, the levels of antibodies generated were too low to be effective despite the use of these harsh adjuvants.
  • Subsequently, Schenk et al. ([0011] Nature, 1999; 400:173-177) showed that immunization with Aβ1-42 peptide inhibits the formation of amyloid plaques and the associated dystrophic neurites in a mouse model of AD. However, due to the low immunogenicity of the Aβ1-42 peptide, the method employed required repeated administrations of the antigen with a harsh lesion-forming adjuvant to obtain the higher levels of anti-Aβ1-42 plaque antibodies necessary to affect plaque formation. Moreover, it was cautioned that immunization with Aβ1-42 might induce more accumulation of the toxic amyloid itself (Araujo, D M & Cotman, C W, Brain Res, 1992; 569, 141-145).
  • Despite these criticisms, additional studies in transgenic AD mice models through similar active immunization have lent credence to the immunoprophylaxis and immunotherapeutic approaches for AD. Janus et al. ([0012] Nature, 2000; 408:979-982) described Aβ1-42 peptide immunization in a mouse model for AD that reduced behavior impairment and plaques. Morgan et al. (Nature, 2000; 408:982-985) described Aβ1-42 peptide vaccination to prevent memory loss in the mouse model.
  • Direct support for the effectiveness of immune therapy came from the observation that peripheral administration of antibodies, monoclonal or polyclonal, against Aβ-peptide reduced amyloid burden (WO 99/27944; Bard et al., [0013] Nature Medicine, 2000; 6:916-919). Despite relatively modest serum levels, these passively administered antibodies, monoclonal 3D6 (anti-Aβ15) and 10D5 (anti-Aβ1-12) or polyclonal anti-Aβ1-42, were able to enter the central nervous system. There, the antibodies bound to plaques and induced clearance of pre-existing amyloid plaques. Bard et al., reported that when examined in an ex vivo assay with brain sections of PDAPP mice (i.e., mice transgenic for an APP mini-gene driven by a platelet-derived growth factor promoter) or AD patient brain tissue, antibodies against Aβ-peptide triggered microglial cells to clear plaques through Fc receptor-mediated phagocytosis and subsequent peptide degradation. This study demonstrated that passively administered antibodies against Aβ1-42 peptide and the Aβ1-42 N-terminus region reduced the extent of plaque deposition in a mouse model of AD; and that monoclonal antibodies or polyclonal antibodies elicited by site-directed vaccines are able to enter the CNS at therapeutically relevant levels.
  • Despite the promising findings of immunological intervention in mice model for AD, a vaccine against AD suitable for humans remains a long way off (Chapman, [0014] Nature, 2000; 408:915-916). The principal hurdles reside in the extensive work necessary to design and formulate an immunogenic composition that is uselful in humans before a practicable vaccine for AD can be achieved. Some of the issues that rely on experimental data for guidance are: (1) What is the specific target site for antibody recognition within the Aβ? (2) In what form should the immunogen be presented? (3) What other sites need to be included before an immunogen is achieved that will elicit a therapeutic level of antibody? (4) What is an effective vaccine delivery system employing a clinically acceptable adjuvant for humans?.
  • A major gap exists between what has been disclosed in the literature and what remains to be done. What is the suitable specific target site (i.e., the polymerized Aβ[0015] 1-42 plaque or the monomeric soluble Aβ1-42 peptide) and how the specific site is to be engineered for immunological intervention. In spite of some 5,000 publications on Aβ1-42 over the past decade, the amyloid cascade hypothesis is hotly debated and the issue: the form in which Aβ1-42 should be used for intervention remains contentious. At the heart of the problem, argued by Terry and colleagues, is the weak correlation between fibrillar amyloid load and measures of neurological dysfunction (The Neuropathology of Alzheimer Disease and the Structure Basis of its Alterations, Ed. by Terry et al., Alzheimer Disease, p187-206, Lippincott Williams and Wilkins, 1999).
  • In AD patients, amyloid deposits often form at a distance from the site of neuron damage. The best correlation with pathological dementia is loss of synaptic terminals. However, the loss of synaptic terminals correlates poorly with amyloid load. If the manifestations of disease correlate weakly with amyloid load, then what is the role of Aβ? The article by Klein et al, titled “Targeting small Aβ[0016] 1-42 oligomers: the solution to an Alzheimer's disease conumdrum?” (Trends in Neurosciences, 2001; 24:219-224) suggests that fibrils are not the only toxic form of Aβ, and perhaps not the neurotoxin that is most relevant to AD. Small oligomers and protofibrils, also termed as Aβ1-42 derived diffusible ligands (ADDLs), may also have potent toxic neurological activity.
  • An AD vaccine for successful immunological intervention will require an immunogen designed to elicit site-directed high affinity antibodies that bind to the senile plaques in the brain tissue to accelerate the clearance of the plaque by the Glial cells, and immunoneutralize the soluble Aβ-derived toxins. [0017]
  • The problem of raising high affinity site-directed antibodies against poorly immunogenic site-specific peptides have been known for decades. Immunologists and vaccinologists often resort to the classical hapten [peptide]-carrier protein conjugate approach as demonstrated in WO 99/27944. For the development of a site-directed vaccine against AD, Frenkel et al. attempted immunization against Aβ[0018] 1-42 plaques through “EFRH”-phage administration (Proc Natl Acad. Sci 2000; 97:11455-11459, WO 01/18169) as mentioned above.
  • The approaches: using Aβ[0019] 1-42 peptide aggregate or Aβ1-42 peptide fragment-carrier protein conjugates (WO99/27944) and using filamentous phage displaying “EFRH peptide” as the agents to induce immune responses against an amyloid deposit in a patient, are cumbersome and ineffective. For example, after the fourth immunization of 1011 phages displaying the EFRH epitope, >95% of the antibodies in the guinea pig immune sera are against the phages. Only a small population (<5%) of the antibodies is against the soluble Aβ1-42 peptide (Frenkel et al., Vaccine 2001,19:2615-2619, WO 01/18169).
  • Less cumbersome methods were described in EP 526,511 and WO 99/27944, which disclosed the administration of Aβ[0020] 1-42 peptide to treat patients with pre-established AD and the administration of Aβ1-42 or other immunogens to a patient under conditions that generate a “beneficial” immune response in the AD patient. However, a review of WO99/27944 show that there are major deficiencies in the vaccine design disclosed therein.
  • In particular, the problem lies in the lack of a pharmaceutically acceptable and effective vaccine delivery system. WO99/27944 disclosed Aβ[0021] 1-42 or active fragments of Aβ1-42 conjugated to a carrier molecule such as cholera toxin as the active vaccine component. See page 4 of WO 99/27944. Although page 5 taught that a pharmaceutical composition comprising the immunogen should be free of Complete Freund's Adjuvant [CFA], the only examples showing the efficacy of the Aβ1-42 vaccine for the treatment of AD in transgenic mice employed large doses of aggregated Aβ1-42 peptide in CFA. Despite repetitive recital of preferred adjuvants that are to be used with the disclosed immunogenic agents to enhance the immune response, experimental data showed that only the formulations employing CFA/ICFA provided a sufficiently high titer of anitbodies. See, page 25 of WO 99/27944. In example 1, the prophylactic efficacy of Aβ1-42 against AD was demonstrated in PDAPP mice. However, the formulations administered consist a dose of 100 ug per mouse of aggregated Aβ1-42 emulsified in Complete Freund's Adjuvant [CFA] (p34 of WO 99/27944) followed by multiple booster doses of the same Aβ1-42 peptide emulsified in Incomplete Freund's Adjuvant. In Example IX, the immune responses in mice to different adjuvants were studied. When the adjuvants: MPL, Alum, QS21, and CFA/ICFA were used with the purportedly potent immunogen AN1792 (i.e., aggregated human Aβ1-42), the level of antibodies to Aβ1-4 were reduced at a statistically significant level in comparison to mice that received the CFA/ICFA vaccines. See, Table 9, and pages 59-64 of WO 99/27944.
  • In the case where Aβ[0022] 1-42 peptide fragments were used (human Aβ1-42 peptides of amino acids 1-5,1-12, 13-28, and 33-42), each fragment was conjugated to sheep anti-mouse IgG as the protein carrier. In a later disclosure, the efficacy of antibodies to Aβ peptide fragments could only be shown by passive immunization with monoclonal antibodies (Bard et al., Nature Medicine 2000; 6:916-919). The efficacy of these fragments conjugated to sheep anti-mouse IgG was not shown. Therefore, the only immunogen shown to be effective was the aggregated Aβ1-42 peptide in CFA/ICFA.
  • Up to the present, all of the vaccine formulations shown to be effective employed CFA/IFA as the adjuvant. Peptide immunogens targeting Aβ[0023] 1-42 have thus far been prepared by conjugation of the various Aβ1-42 fragments to sheep anti-mouse immunoglobulin, conjugation of synthetic Aβ13-28 via m-maleimidobenzoyl-N-hydroxysuccinimide ester to anti-CD3 antibody, or aggregated Aβ1-42 peptide alone. These immunogens, i.e., Aβ1-42 peptide alone or Aβ1-42 peptide-carrier protein conjugates, were emulsified with complete Freund's adjuvant for the first immunization, followed by subsequent boosts in incomplete Freund's adjuvant (Johnson-Wood et al., Proc Natl Acad Sci USA, 1997; 94:1550-1555; Seubert et al., Nature, 1992; 359:325-327; Schenk et al., Nature, 1999; 400: 173-177; Janus et al., Nature 2000; 408:979-982; and Morgan et al., Nature, 2000; 408:982-985). The formulations disclosed in WO 99/27944 or others using CFA and ICFA as adjuvants causes lesions and are too harsh for use in humans. Thus, none of the vaccine compositions for AD described in the prior art are suitable for use in humans.
  • In summary, despite statements suggesting the potential of Aβ[0024] 1-42 peptide for the treatment of AD in view of the previous disclosures of Kline (EP 526,511), no problem solving vaccine formulations were really offered in WO99/27944 to address this key problem.
  • Another disadvantage with the peptide-carrier protein conjugates and Aβ[0025] 1-42 aggregates is that these molecules are highly complex and are difficult to characterize and it is difficult to develop effective quality control procedures for the manufacturing process. A further disadvantage is that, Aβ1-42 peptide or its fragments are self molecules when administered to humans. Therefore, they are less immunogenic or non-immunogenic in humans. It is, thus, necessary to develop clinically acceptable vaccine formulations for administration in humans.
  • It is known that promiscuous Th epitopes may be employed to evoke efficient T cell help and may be combined with poorly immunogenic B cell epitopes to provide potent immunogens. Well-designed promiscuous Th/B cell epitopes chimeric peptides have been shown to be useful in eliciting Th responses and resultant antibody responses in most members of a genetically diverse population expressing diverse MHC haplotypes. Promiscuous Th from a number of pathogens, such as measles virus F protein and hepatitis B virus surface antigen, are known. Tables 1 and 2 lists many of the known promiscuous Th that have been shown to be effective in potentiating a short poorly immunogenic peptide, the decapeptide hormone LHRH (U.S. Pat. No. 5,759,551, and 6,025,468). [0026]
  • Potent Th epitopes range in size from approximately 15-40 amino acid residues in length, often share common structural features, and may contain specific landmark sequences. For example, a common feature of a Th is that it contains amphipathic helices, alpha-helical structures with hydrophobic amino acid residues dominating one face of the helix and with charged and polar residues dominating the surrounding faces (Cease et al., [0027] Proc Natl Acad Sci USA, 1987; 84: 4249-4253). Th epitopes frequently contain additional primary amino acid patterns such as a Gly or charged residue followed by two to three hydrophobic residues, followed in turn by a charged or polar residue. This pattern defines what are called Rothbard sequences. Th epitopes often obey the 1, 4, 5, 8 rule, where a positively charged residue is followed by hydrophobic residues at the fourth, fifth and eighth positions after the charged residue. Since all of these structures are composed of common hydrophobic, charged and polar amino acids, each structure can exist simultaneously within a single Th epitope (Partidos et al., J Gen Virol, 1991; 72:1293). Most, if not all, of the promiscuous T cell epitopes fit at least one of the periodicities described above. These features may be incorporated into the designs of idealized artificial Th sites, including combinatorial Th epitopes. With respect to the design of combinatorial Th sites, lists of variable positions and preferred amino acids are available for MHC-binding motifs (Meister et al., Vaccine, 1995; 13:581-591). Furthermore, a method for producing combinatorial Th has been disclosed for combinatorial library peptides termed structured synthetic antigen library (Wang et al., WO 95/11998). Thus, the 1, 4, 5, 8 rule can be applied together with known combinatorial MHC-binding motifs to assign invariant and degenerate positions in a combinatorial Th site, and to select residues for the degenerate sites to vastly enlarge the range of immune responsiveness of an artificial Th. See, Table 2, WO 99/66957, and WO 95/11998.
  • Wang et al. (U.S. Pat. No. 5,759,551) suggested the use of immunostimulatory elements to render the self protein Amylin immunogenic. Wang et al. suggested the administration of immunogenic synthetic amylin peptides as vaccines for the treatment of non-insulin dependent diabetes mellitus (NIDDM), an amyloidogenic disease caused by overproduction of Amylin (column 19, lines 9-39, U.S. Pat. No. 5,759,551). Amylin is a 37 amino acid residues peptide hormone produced by the β cells in the islets of Langerhans. Overproduction of Amylin will result in the depositon of insoluble amyloid leading to amyloidogenic disease in the pancreas. Similar to the overproduction of Amylin, overproduction of the Aβ[0028] 1-42 peptide will lead to the deposition of insoluble amyloid in the brain of AD patients. However, there is limited sequence homology between Amylin and the Aβ1-42 peptide. Only a short stretch of amino acids residues, VGSN, of Amylin32-35 corresponds to Aβ24-27. Antibodies produced against the Amylin peptide is not expected to be cross reactive to soluble Aβ1-42 peptides nor accelerate the clearance of amyloid plaques in the brain in view of the studies by Soloman et al. and Schenk et al., which showed that the sequence EFRH is critical.
  • It is the object of the invention to develop an immunogen that will enable the generation of high levels of high affinity antibodies against the N-terminal functional site of the Aβ[0029] 1-42 peptide with high cross-reactivity to the senile plaques in the brain of AD patients. The anitbodies generated by binding to the Aβ1-42 peptide and the senile plaques is expected to accelerate the clearance of these plaques from the brain, promote fibril disaggregation, inhibit fibrillar aggregation, and immunoneutralization of the “soluble Aβ-derived toxins” [also termed as Aβ-derived diffusible ligands or ADDLs].
  • It is a further objective of the present invention to develop a vaccine delivery vehicle that is suitable for human or veterinary use for the prophylaxis and treatment of Alzheimer's Disease.[0030]
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • FIGS. 1[0031] a, 1 b, 1 c, 1 d, 1 e and If are photographs showing Immunoperoxidase staining of serial sections from 2 AD brains, using Avidin-Biotinylated Antibody Complex (ABC) method with immune and preimmune sera at 1:100 dilution under 10× magnification. FIGS. 1a, and 1 d show significant binding of antibodies to both senile plaques and Aβ plaques (both labelled as “P”) on thioflavine S positive blood vessels (labelled as “BV”). The antibodies were generated in guinea pigs using Aβ1-28-εK-MVF Th1-16 (SEQ ID NO:74) prepared in ISA51 water-in-oil emulsion. FIGS. 1b and 1 e show the cross reactivity of antibodies raised against the same peptide immunogen in CFA/ICFA. FIGS. 1c and 1 f show brain sections using preimmune sera.
  • FIGS. 2[0032] a, 2 b, 2 c, 2 d, and 2 e are photographs showing Immunoperoxidase staining of serial sections of AD brain with immune and preimmune sera at 1:100 dilution and under 40× magnification. FIGS. 2a and 2 d showed that the antibodies in guinea pigs immunized with Aβ1-28-εK-MVF Th-1-16 (SEQ ID NO:74) prepared in ISA51 water-in-oil emulsion strongly stained the plaques (P) forming a pattern of cores. FIG. 2b is a photograph of the staining pattern of AD pig brain sections using the same immunogen in CFA/ICFA formulation. The anti-sera reacted predominantly with plaques on the blood vessels (BV). FIG. 2c is a photograph of a guinea pig brain section with preimmune serum and showed no staining. FIG. 2e shows the brain section with hyperimmune sera generated by immunization with Aβ1-28 peptide alone in CFA/ICFA showing a surprisingly weak staining pattern despite the strong reactivity with Aβ1-28 by ELISA.
    • BRIEF DESCRIPTION OF THE INVENTION
  • The present invention relates to an immunogenic composition comprising synthetic peptides capable of inducing antibodies against the main functional/regulatory site of the Aβ peptide with high cross-reactivity both to the soluble Aβ[0033] 1-42 peptide and the plaques in the brain of Alzheimer's Disease (AD) patients. The immunogenic composition when administered to an AD patient or a person predisposed to AD is expected to accelerate the clearance of amyloid plaques and immunoneutralization of the soluble Aβ derived toxins in the brain to prevent and treat AD. In particular, a peptide immunogen of this invention comprise a Th epitope selected from the group consisting of SEQ ID NOS: 1-64 and the immunologically functional analogs thereof linked to a short N-terminal Aβ1-42 peptide fragment selected from the group consisting of 10 to 28 amino acid residues comprising EFRH of the Aβ1-42 peptide, SEQ ID NO:65, or an immunologically functional analog of the Aβ1-42 peptide fragment. Preferably the Aβ1-42 peptide fragment is selected from the group SEQ ID NOS: 66-69 or a immunologically functional analogs thereof.
  • The present invention further provides an immunogenic composition comprising an immunologically effective amount of a peptide composition in a pharmaceutically acceptable vaccine formulation comprising an adjuvant or emulsifier selected from the group consisting of liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montanide ISA51, ISA35, ISA206 and ISA 720 as well as the other efficacious adjuvants and emulsifiers. [0034]
  • The present invention further provides a method for the induction of accelerated clearance of amyloid plaques and immunoneutralization of the soluble Aβ-derived toxins in the brain to prevent and treat Alzheimer Disease in a mammal by administering one or more of the immunogenic peptides to the mammal for a time and under conditions sufficient to induce antibodies directed against the functional/regulatory site of the Aβ[0035] 1-42 peptide. A typical example of a vaccine of the present invention is a peptide composition comprising 5-1000 μg of the peptide immunogen in a vaccine formulated as a water in oil emulsion in a pharmaceutically acceptable adjuvant and/or carrier. A typical method of administering the vaccine is to inject intramuscularly the vaccine formulation at 0.5-2 mL per dose on an immunization schedule of 0, 4, and 8 weeks intervals.
  • Yet another aspect of the invention relates to an immunogenic synthetic peptide of about 30 to about 60 amino acids consisting of a helper T cell (Th) epitope, linked to an N-terminal Aβ[0036] 1-42 peptide fragment selected from the group consisting of 10 to 28 amino acids with each fragment comprising amino acid residue 1 of the Aβ1-42 peptide. See SEQ ID NO:65 wherein D, Aspartic acid, is designated as amino acid residue 1. Preferably the N terminal Aβ1-42 peptide fragment is selected from the group SEQ ID NOs: 66-69 or a peptide analog of N-terminal fragment of Aβ1-42 peptide. Optionally, amino acid spacers to separate the immunogenic domains may be included. The immunogenic domain elements separated by spacers can be covalently joined in any order provided that either the immunoreactivity of the peptide hapten is substantially preserved or that immunoreactivity to the N-terminal Aβ peptide fragment, soluble Aβ1-42 peptide, and the plaques is generated.
  • An important factor affecting immunogenicity of a synthetic peptide for an N-terminal Aβ[0037] 1-42 fragment immunogen is its presentation to the immune system by T helper cell epitopes (Th). Such Th is most reliably supplied to the peptide immunogen by foreign Th epitopes placed on a separate Th peptide domain element that are extrinsic to the target Aβ peptide. Such peptide immunogens may be produced as hybrid polypeptides by recombinant DNA expression. They may also be more simply and less expensively supplied as a synthetic peptide immunogen comprising the target hapten B cell site from Aβ peptide and T-helper epitopes (Th) appropriate for the host. Such peptides react with helper T-cell receptors and the class II MHC molecules, in addition to antibody binding sites (Babbitt et al., Nature, 1985; 317:359) and thus stimulate a tightly site-specific antibody response to the target antibody binding site. Previously such Th was supplied for workable Aβ1-42 peptide immunogens by Th intrinsic to aggregated full length Aβ peptide (WO 99/66957; WO 1999/27944; Janus et al., 2000, Morgan et al., 2000) and can be supplied by carrier protein. A wholly synthetic peptide immunogen enjoys the following advantages over Aβ1-42 peptide aggregates, carrier conjugates and recombinant polypeptides in that the product is chemically defined for easy quality control. The synthetic peptide immunogen is stable. No elaborate downstream processing nor an elaborate manufacturing facility is needed. The immune response is site-specific and focused on the Aβ target and not the carrier. Thus, undesirable responses such as epitopic suppression are avoided.
  • Immunogenicity of synthetic N-terminal functional-site directed Aβ peptide immunogens can be optimized by (1) combining N-terminal Aβ[0038] 1-42 peptide fragment with selected foreign promiscuous Th sites to which the majority of a population are responsive; and (2) combining Aβ peptide fragment with Th whose repertoire is enlarged through combinatorial chemistry, and thereby accommodate to the variable immune responsiveness of a genetically diverse population.
  • It has been found that peptides composition of the present invention are effective in stimulating the production of antibodies against the main functional/regulatory site of the Aβ peptide, with cross-reactivities to the soluble Aβ[0039] 1-42 and the plaques in the brains of AD patients. Based on the immunogenicity data obtained in guinea pigs and baboons, and the data obtained from the immunoperoxidase staining of the amyloid plaques present in human AD brain sections by the spicific immune sera obtained, it is expected that the peptide immunogens of the present invention formulated appropriately are effective in humans. It is to be noted that the data obtained in baboons are particularly significant in that this is a species whose immune response closely resemble those of humans.
    • DETAILED DESCRIPTION OF THE INVENTION
  • This invention is directed to a novel peptide composition for the generation of high titer polyclonal antibodies with specificity for the main functional/regulatory site of the Aβ peptide, with cross-reactivities to the soluble Aβ[0040] 1-42 and the plaques in the brain of Alzheimer Disease (AD) patients. The antibodies generated by the peptide composition are highly site-specific and bind to the Aβ peptides and to amyloids plaques in the brain. Thus, the present invention provides an effective method for accelerating the clearance of amyloid plaques and immunoneutralization of soluble Aβ derived toxins in the brains for the prevention and treatment of AD.
  • N-terminal Aβ[0041] 1-42 peptide fragments selected from the group consisting of 10 to 28 amino acids wherein each fragment comprises EFRH of the Aβ1-42 peptide (SEQ ID NO:65), are short linear peptides which, by themselves are non-immunogenic. The short Aβ1-42 peptide fragments can be immuno-potentiated by chemical coupling to a carrier protein, for example, keyhole limpet hemocyanin (KLH) or by fusion to a carrier polypeptide through recombinant DNA expression, for example, hepatitis B surface antigen. The deficiency of such “Aβ peptide(s)-carrier protein” vaccines is that a major portion of antibodies generated are non-functional antibodies directed against the carrier protein.
  • The immunogens of the present invention are wholly synthetic peptide immunogens comprising N-terminal fragment of Aβ[0042] 1-42 peptide of 10 to 28 amino acids with each fragment comprising EFRH of the Aβ1-42 peptide covalently linked to promiscuous Th epitopes selected from the group consisting of SEQ ID NOs: 1 to 64. The immunogens of the invention elicit the production of site-specific antibodies which bind to the Aβ1-42 peptide and its aggregates and are cross reactive with amyloid plaques in the brain to provide for accelerated clearance of amyloid plaques and immunoneutralization of the soluble Aβ-derived toxins in the brain. Thus, the immunogen of the present invention is useful in preventing and treating AD.
  • The helper T cell epitopes (Th) useful in the invention comprise multiple class II MHC binding motifs. Specific examples of Th covalently linked to an N-terminal Aβ[0043] 1-42 peptide fragment are provided. The results of anti-sera from animals immunized with the immunogen peptides of the present invention demonstrate that potent site-directed Aβ peptide reactive antibodies are generated, in a genetically diverse host population.
  • Generally, the synthetic immunogenic peptide of the present invention are approximately 20 to 100 amino acids long and comprise: [0044]
  • (i) a helper T cell (Th) epitope selected from the group consisting of SEQ ID Nos: 1 to 64; [0045]
  • (ii) an N-terminal fragment of Aβ[0046] 1-42 peptide from about 10 to about 28 amino acid residues wherein each fragment comprises EFRH of the Aβ1-42 peptide; and
  • (iii) optionally a spacer consisting of at least an amino acid to separate the immunogenic domains. [0047]
  • Preferably, the N terminal fragment of the Aβ[0048] 1-42 peptide is selected from the group consisting of SEQ ID NOS: 66-69 and an immunologically effective analog thereof. The Th peptide is covalently attached to either the N— or C-terminus of the target N-terminal fragment of Aβ1-42 peptide optionally with a spacer (e.g., Gly-Gly, ε-N Lys).
  • The peptide immunogen of this invention is represented by one of the following formula: [0049]
  • (A)n-(N-terminal fragment of Aβ1-42 peptide)-(B)o-(Th)m-X; or
  • (A)n-(Th)m-(B)o-(N-terminal fragment of Aβ1-42 peptide)-X;
  • wherein [0050]
  • each A is independently an amino acid; [0051]
  • each B is a linking group selected from the group consisting of an amino acid, gly-gly, (α, ε-N)lys, Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:73); [0052]
  • Each Th comprise an amino acid sequence that constitutes a helper T cell epitope, or an immune enhancing analog or segment thereof; [0053]
  • (N-terminal fragment of Aβ[0054] 1-42 peptide) is a synthetic peptide B cell target site antigen and is a fragment of about 10 to about 28 amino acid residues wherein each fragment comprises EFRH of the Aβ1-42 peptide or an immunologically functional analog thereof;
  • X is an α-COOH or α-CONH[0055] 2 of an amino acid;
  • n is from 0 to about 10; [0056]
  • m is from 1 to about 4; and [0057]
  • o is from 0 to about 10. [0058]
  • The peptide immunogen of the present invention comprises from about 20 to about 100 amino acid residues, preferably from about 25 to about 60 amino acid residues. Preferably, the (N-terminal fragment of Aβ[0059] 1-42 peptide) is selected from the group consisting of SEQ ID Nos: 66-69 and preferably the Th epitope is selected from the group consisting of SEQ ID NOs: 1, 3, 4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54. Preferably, m=1, n=1, and o=1 or 2.
  • When A is an amino acid, it is a non-naturally occurring or naturally occurring amino acid. Non-naturally occurring amino acids include, but are not limited to, ε-N lysine, β-alanine, ornithine, norleucine, norvaline, hydroxyproline, thyroxine, γ-amino butyric acid, homoserine, citrulline and the like. Naturally-occurring amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine. when m is greater than one, and two or more of A are amino acids, then each amino acid may independently be the same or different. (A)[0060] n may include a spacer, e.g., Gly-Gly, ε-N Lys.
  • B is a spacer and is an amino acid which can be naturally occurring or the non-naturally occurring amino acids as described above. Each B is independently the same or different. The amino acids of B can also provide a spacer, e.g., Gly-Gly, ε-Lys, or lysine between the promiscuous Th epitope and the N-terminal fragment of Aβ[0061] 1-42 peptide (e.g., SEQ ID NOs:66-69) or an immunologically functional analog thereof. In addition by physically separating the Th epitope from the B cell epitope, i.e., the N-terminal fragments of Aβ1-42 peptide or its immunologically functional analog, the Gly-Gly or F-Lys spacer can disrupt any artifactual secondary structures created by the joining of the Th epitope with an N-terminal fragment of Aβ1-42 peptide or its immunologically functional analog and thereby eliminate interference between the Th and/or B cell responses. The amino acids of B can also form a spacer which acts as a flexible hinge that enhances separation of the Th and the N-terminal fragments of Aβ1-42 peptide. Examples of sequences encoding flexible hinges are found in the immunoglobulin heavy chain hinge region. Flexible hinge sequences are often proline rich. One particularly useful flexible hinge is provided by the sequence Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:77), where Xaa is any amino acid, and preferably aspartic acid. The conformational separation provided by the amino acids of B permits more efficient interactions between the presented peptide immunogen and the appropriate Th cells and B cells to enhances the immune responses to the Th epitope and the antibody-eliciting epitope or their immunologically functional analogs.
  • Th is a sequence of amino acids (natural or non-natural amino acids) that comprises a Th epitope. A Th epitope may be a continuous or discontinuous epitope. In a discontinuous Th epitope, not every amino acid of Th is necessary. A Th epitope, or an analog or fragment thereof, is capable of enhancing or stimulating an immune response to the N-terminal fragment of Aβ[0062] 1-42 peptide. Th epitopes that are immunodominant and promiscuous are highly and broadly reactive across animal and human populations with widely divergent MHC types (Partidos et al., 1991; U.S. Pat. No. 5,759,551). The Th epitope of the subject peptides is about 10 to about 50 amino acids, preferably from about 10 to about 30 amino acids. When multiple Th epitopes are present (i.e., m>2), each Th epitope may be the same or different. A Th segment comprises a contiguous portion of a Th epitope that is sufficient to enhance or stimulate an immune response to the N-terminal fragment of Aβ1-42 peptide.
  • Th epitopes of the present invention include those derived from foreign pathogens including but not limited to those exemplified in Table 1 (SEQ ID Nos:1-21). Further, Th epitopes include idealized artificial Th and artificial idealized combinatorial Th disclosed in WO 99/66957 and listed here in Table 2 as SEQ ID Nos 22-64. Peptides comprising combinatorial Th are produced simultaneously in a single solid-phase peptide synthesis in tandem with the N-terminal fragment of Aβ[0063] 1-42 peptide, A and B. The Th epitopes also include immunologically functional analogs thereof, having conservative substitutions, additions, deletions and insertions therein of from one to about 10 amino acid residues as long as the Th-stimulating function has not been essentially modified.
  • In the synthetic peptides of this invention, the Th epitope is covalently attached through a spacer B to either the N terminus or C terminus of the N-terminal fragment of Aβ[0064] 1-42 peptide or an immunologically functional analog thereof. An immunologically functional analog of the N-terminal fragment of Aβ1-42 peptide may comprise conservative substitutions, additions, deletions, or insertions of from one to about four amino acid residues as long as immune responses that are crossreactive with the Aβ1-42 peptides are elicited. The conservative substitutions, additions, and insertions can be accomplished with natural or non-natural amino acids as defined above.
  • The preferred peptide immunogens of this invention are those comprising the N-terminal fragment of the Aβ[0065] 1-42 peptide fragments selected from the group consisting of SEQ ID NOs: 66-69 or an immunologically functional analog thereof; a spacer (e.g., Gly-Gly, ε-Lys); a Th epitope selected from the group consisting of an HBs Th (SEQ ID NO:1); HBc Th (SEQ ID NO:20); MVF Th (SEQ ID NOS:8, 9); PT Th (SEQ ID NOs:4, 5, 7), TT Th (SEQ ID NOs:3, 4, 6); CT Th (SEQ ID NOs:12, 21); DT Th (SEQ ID NO:13, 14), MVF Th derived artificial Th selected from the group consisting of SEQ ID Nos:38-40, 47-51); HBV Th derived artificial Th selected from the group consisting of SEQ ID NOS: 52-54. See Tables 1 and 2.
  • Peptide compositions which contain a cocktail of the subject peptide immunogens with two or more Th epitopes may enhance immunoefficacy in a broader population and thus provide an improved immune response to the Aβ[0066] 1-42 peptides and their fragments.
  • The peptide immunogens of this invention can be made by chemical synthesis methods which are well known to the ordinarily skilled artisan. See, for example, Fields et al., Chapter 3 in [0067] Synthetic Peptides: A User's Guide, ed. Grant, W. H. Freeman & Co., New York, N.Y., 1992, p. 77. Hence, peptides can be synthesized using the automated Merrifield techniques of solid phase synthesis with the α-NH2 protected by either t-Boc or F-moc chemistry using side chain protected amino acids on, for example, an Applied Biosystems Peptide Synthesizer Model 430A or 431. Preparation of peptide constructs comprising combinatorial library peptides for Th epitopes can be accomplished by providing a mixture of alternative amino acids for coupling at a given variable position. After complete assembly of the desired peptide immunogen, the resin is treated according to standard procedures to cleave the peptide from the resin and deblock the functional groups on the amino acid side chains. The free peptide is purified by HPLC and characterized biochemically, for example, by amino acid analysis or by sequencing. Purification and characterization methods for peptides are well known to one of ordinary skill in the art.
  • The immunogen of the present invention may also be prepared as a branched polymer by synthesis of the desired peptide construct directly onto a branched poly-lysyl core resin (Wang, et al., [0068] Science, 1991; 254:285-288).
  • Alternatively, the longer synthetic peptide immunogens can be synthesized by well known recombinant DNA techniques. Such techniques are provided in well-known standard manuals with detailed protocols. To construct a gene encoding a peptide of this invention, the amino acid sequence is reverse translated to obtain a nucleic acid sequence encoding the amino acid sequence, preferably with codons that are optimum for the organism in which the gene is to be expressed. Next, a synthetic gene is made, typically by synthesizing oligonucleotides which encode the peptide and any regulatory elements, if necessary. The synthetic gene is inserted in a suitable cloning vector and transfected into a host cell. The peptide is then expressed under suitable conditions appropriate for the selected expression system and host. The peptide is purified and characterized by standard methods. [0069]
  • The efficacy of the peptide composition of the present invention can be established by injecting an animal, for example, guinea pigs, with an immunogenic composition comprising peptides of the invention. See, Table 4, SEQ ID NOS:70-75. The humoral immune response to the N-terminal fragment of Aβ[0070] 1-42 peptide and the soluble Aβ1-42 peptide are monitored. A detailed description of the procedures used is provided in the Examples hereinbelow.
  • Another aspect of this invention provides a peptide composition comprising an immunologically effective amount of one or more of the peptide immunogens of this invention in a pharmaceutically acceptable delivery system. Accordingly, the subject peptide composition can be formulated as a vaccine using pharmaceutically acceptable adjuvants, carriers or other ingredients routinely employed in the formulation of vaccines. Among the ingredients that can be used in this invention are adjuvants or emulsifiers including alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720 as well as the other efficacious adjuvants and emulsifiers. The composition may be formulated for immediate release or sustained release. The composition may also be formulated for induction of systemic immunity, e.g., by entrapment in or coadministration with microparticles. Such formulations are readily available to one of ordinary skill in the art. [0071]
  • The immunogens of the present invention can be administered via any conventional route, such as subcutaneous, oral, intramuscular, parenteral or enteral route. The immunogens can be administered in a single dose or in multiple doses. A suitable immunization schedule is readily determined and available to one of ordinary skill in the art. [0072]
  • The peptide composition of the present invention comprises an effective amount of one or more of the peptide immunogens of the present invention and a pharmaceutically acceptable carrier. Such a composition in a suitable dosage unit form generally contains about 0.25 μg to about 500 μg of the immunogen per kg body weight. When delivered in multiple doses, the effective amount may be conveniently divided per dosage unit. For example, an initial dose, e.g. 0.0025-0.5 mg per kg body weight; preferably 1-50 μg per kg of body weight of the peptide immunogen is to be administered by injection, preferably intramuscularly, followed by repeat (booster) doses of a similar amount. Dosage will depend on the age, weight and general health of the subject as is well known in the vaccine and therapeutic arts. [0073]
  • The immune response of the synthetic Aβ[0074] 1-42 peptide immunogens can be improved by delivery through entrapment in or on biodegradable microparticles of the type described by O'Hagan et al. (Vaccine, 1991; 9: 768-771). The immunogens can be encapsulated with or without an adjuvant in biodegradable microparticles, to potentiate immune responses, and to provide time-controlled release for sustained or periodic responses, and for oral administration, (O'Hagan et al., 1991; and, Eldridge et al., 1991; 28: 287-294).
  • The following examples are provided to illustrate the invention. The scope of the invention is not to be limited to the specific peptide immunogens and compositions provided. The examples demonstrate that the peptide immunogens of the present invention are useful for eliciting site-directed antibodies to both Aβ[0075] 1-10 and Aβ1-14 fragments as well as cross-reactive antibodies to soluble Aβ1-42 peptides as early as 4 weeks after the initial immunization.
    • EXAMPLE 1 Typical Methods to Synthesize Aβ Peptide Immunogens of the Present Invention
  • Peptide immunogens listed in Table 4 (SEQ ID NOS:70-76) were synthesized individually by the Merrifield solid-phase synthesis technique on Applied Biosystems automated peptide synthesizers (Models 430, 431 and 433A) using Fmoc chemistry. Preparation of peptide immunogens comprising a combinatorial library Th, i.e., idealized artificial Th site such as MvF derived Th1-8 (SEQ ID NOs:38-40), can be accomplished by providing a mixture of the desired amino acids for chemical coupling at a given position as specified in the design. After complete assembly of the desired peptide, the resin was treated according to standard procedure using trifluoroacetic acid to cleave the peptide from the resin and deblock the protecting groups on the amino acid side chains. The cleaved, extracted and washed peptides were purified by HPLC and characterized by mass spectrometry and reverse phase HPLC. [0076]
    • EXAMPLE 2 Evaluation of the Immunogenicity of the Aβ Peptide Immunogens of the Present Invention
  • Aβ-derived peptide immunogens were evaluated on groups of guinea pigs as specified by the experimental immunization protocol outlined below and by serological assays for determination of immunogenicity. [0077]
  • Standard Experimental Design: [0078]
  • Immunogens: [0079]
  • (1) individual peptide immunogen; or [0080]
  • (2) a mixture of equal molar peptide immunogens as specified in each example. [0081]
  • Dose: 100 μg in 0.5 mL per immunization unless otherwise specified [0082]
  • Route: intramuscular unless otherwise specified [0083]
  • Adjuvants: Complete Freund's Adjuvant (CFA)/Incomplete [0084]
  • Adjuvant (IFA); or water in oil emulsions unless otherwise specified. [0085]
  • CFA/IFA groups received CFA week 0, IFA in subsequent weeks. [0086]
  • Dose Schedule: 0, 3, and 6 weeks or otherwise specified. [0087]
  • Bleed Schedule: weeks 0, 5, 8 or otherwise specified [0088]
  • Species: Duncan-Hartley guinea pigs or otherwise specified [0089]
  • Assay: Specific ELISAs for each immune serum's anti-peptide activity. The Solid phase substrate was the Aβ peptide fragment e.g. Aβ[0090] 1-14 or full length Aβ1-42 (SEQ ID NOs: 67 and 65). Blood was collected and processed into serum, and stored prior to ELISA with the target peptides.
  • The immunoreactivities of the antibodies elicited against Aβ peptides and against the soluble Aβ[0091] 1-42 peptides were determined by ELISAs (enzyme-linked immunosorbent assays) using 96-well flat bottom microtiter plates which were coated with the Aβ1-42 peptide fragments, SEQ ID NOs: 67 or 65 as the immunosorbent. Aliquots (100 μL) of the peptide immunogen solution at a concentration of 5 μg/mL were incubated for 1 hour at 37° C. The plates were blocked by another incubation at 37° C. for 1 hour with a 3% gelatin/PBS solution. The blocked plates were then dried and used for the assay. Aliquots (100 μL) of the test immune sera, starting with a 1:100 dilution in a sample dilution buffer and ten-fold serial dilutions thereafter, were added to the peptide coated plates. The plates were incubated for 1 hour at 37° C.
  • The plates were washed six times with 0.05% PBS/Tween® buffer. 100 μL of horseradish peroxidase labeled goat-anti-species specific antibody was added at appropriate dilutions in conjugate dilution buffer (Phosphate buffer containing 0.5M NaCl, and normal goat serum). The plates were incubated for 1 hour at 37° C. before being washed as above. Aliquots (100 μL) of o-phenylenediamine substrate solution were then added. The color was allowed to develop for 5-15 minutes before the enzymatic color reaction was stopped by the addition of 50 μL 2N H[0092] 2SO4. The A492 nm of the contents of each well was read in a plate reader. ELISA titers were calculated based on linear regression analysis of the absorbances, with cutoff A492 nm set at 0.5. The cutoff value chosen was rigorous with the values for diluted normal control samples being less than 0.15.
    • EXAMPLE 3 Characterization of the Relative Immunogenicities of Aβ1-42 and its N-Terminal Fragments for Optimization of Design For Site-Directed Aβ Peptide-Based Synthetic Vaccine
  • To design a total synthetic vaccine that generates a high level of high affinity antibodies against the Aβ peptides with high cross-reactivity to the soluble Aβ[0093] 1-42 peptides and the plaques in the brain of AD patients, the relative immunogenicities of Aβ1-42 and its N-terminal fragments were characterized initially. In order to determine the relative immunological properties of the various regions within Aβ1-42 peptide, a mild adjuvant suitable for human use, alum was employed in the first study. The relative immunogenicities of Aβ1-42 peptide and an N-terminal fragment thereof, Aβ1-28 were compared. The immunogenicity evaluation was conducted according to procedures described in Example 2. Unexpectedly, Aβ1-28 was found to be more immunogenic than the Aβ1-42 peptide, indicating that there is immunosuppression within C-terminal fragment Aβ29-42 (Table 5).
  • Subsequently, the immunogenicities of Aβ[0094] 1-28 was compared to Aβ1-14, a shorter N-terminal fragment of Aβ1-42. A more potent adjuvant suitable for human use (Montamide ISA51, Seppic, Paris, FR) was employed for the preparation of a water-in-oil emulsion for formulating the vaccine. Based on the data obtained as shown in Table 6, the relative immunogenicities for the three Aβ peptides (i.e. Aβ1-14, Aβ1-28 and Aβ1-42) were ranked Aβ1-28>Aβ1-42>Aβ1-14. Surprisingly, the loss of the C-terminal 14 mer from Aβ1-42, improved rather than reduced the immunogenicity. The antibody response against Aβ is primarily directed to the N-terminal region, particularly the Aβ1-14 N-terminal fragment as shown by ELISA data (Table 6). However, a further shortening of the Aβ1-28 fragment from the C-terminal to form the Aβ1-14 fragment resulted in a loss in immunogenicity.
  • The short Aβ[0095] 1-14 fragment contains the main functional/regulatory site, EFRH, located at positions 3-6 of the Aβ1-42 peptide as reported by Solomon et al. The blocking of this epitope by antibodies modulates the dynamics of aggregation as well resolubilization of already formed aggregates (Soloman et al., Proc Nat/Acad. Sci, 1996; 93:452-455; Proc Natl Aca. Sci, 1997; 94:4109-4112). Most of the anti-Aβ1-28 and Aβ1-42 antibodies are directed against the N-terminal fragment of the Aβ1-42 peptide containing this epitope (Table 6). However, the Aβ1-14 fragment by itself was poorly immunogenic. The results of this experiment suggest the presence of an intrinsic Th epitope within the Aβ15-28 segment. This intrinsic Th epitope accounts for the modest immunogenicities of Aβ1-28 and Aβ1-42 peptides in guinea pigs.
  • The presence of a Th epitope in the Aβ[0096] 15-28 fragment is desirable. However, it is desirable to be able to engineer a more potent immunogen for a successful human vaccine when faced with the limitation of a restricted human MHC molecule, the number of appropriate doses and the type of adjuvants permitted for human use. Therefore, we attempted the linkage of a foreign or extrinsic Th such as that derived from HBV Th (SEQ ID NO: 1) to the C-terminal of the Aβ1-28 peptide (SEQ ID NO:66). The extrinsic Th epitope significantly enhanced the immunogenicity of the Aβ1-28 fragment as shown in Table 6. The antibody response to the engineered immunogen with the Aβ1-28 fragment remained directed to the functional N-terminal fragment of peptide immunogen (SEQ ID NO: 70) making this construct a better immunogen than the Aβ1-28 fragment or Aβ1-42 fragment alone. This peptide immunogen (SEQ ID NO: 70) represents a peptide immunogen with the formula:
  • (A)n (N-terminal fragment of Aβ peptide)-(B)o-(Th)m
  • wherein: [0097]
  • A is αNH[0098] 2, with Aβ1-28 being an N-terminal fragment of Aβ1-42;
  • B is glycine; [0099]
  • Th is a helper T cell epitope derived from a foreign pathogen, [0100]
  • HBsAg Th (SEQ ID NO: 1), and wherein n is 1, m is 1 and o is 2. [0101]
    • EXAMPLE 4 Lower Limit of N-Terminal Fragment of Aβ for the Development of Aβ Based Synthetic Vaccine for Ad
  • Since the main functional/regulatory site comprising the EFRH residues is located at positions 3-6 of the Aβ[0102] 1-42 peptide (Soloman et al. Proc Natl Acad. Sci, 1996; 93:452-455; Proc Natl Aca. Sci, 1997; 94:4109-4112), it was useful to explore the shortest N-terminal fragment of Aβ1-42 peptide as an optimal B cell target site on Aβ for incorporation into the synthetic immunogen of the present invention.
  • Each of several short non-immunogenic N-terminal fragments of Aβ, Aβ[0103] 1-10, Aβ1-12, Aβ1-14 along with Aβ1-28 was incorporated into immunogens designed with a representative idealized artificial Th (SEQ ID NO:51). Linkage was through an εN-Lys spacer. The engineered constructs were formulated with strong adjuvants due to the expected low immunogenicity of the short Aβ fragments. The three synthetic constructs were formulated in complete and incomplete Freund's adjuvant and tested for their immunogenicities based on procedures as described in Example 2. As shown in Table 7, all four peptide immunogens were highly immunogenic with Log10 ELISA titers in the range from 4.3 to 5.6 [i.e. 104.3 to 105.6] with very high crossreactivities to the full length Aβ1-42 peptide after only four weeks from the initial immunization. More importantly, fragments as small as Aβ10, Aβ1-12 and Aβ1-14 each linked to the idealized artificial Th (SEQ ID NO:51) were found to be highly immunogenic after linkage to a disclosed artificial Th epitope (Table 7). These peptide immunogens were designed in accordance with the formula:
  • (A)n—(N-terminal fragment of Aβ peptide)-(B)o-(Th)m
  • wherein: [0104]
  • A is αNH[0105] 2, wherein the N-terminal fragment is Aβ1-10, Aβ1-12, Aβ1-14 or Aβ1-28;
  • B is ε-N Lysine, a spacer linked through its epsilon amino group to the next amino acid; [0106]
  • Th is a helper T cell epitope derived from an idealized artificial Th, MVF Th1-16(SEQ ID NO:51), wherein n is 1, m is 1 and o is 1. [0107]
  • It was found that further reduction in the length of the N-terminal fragment of Aβ to less than a 10 mer would result in more limited, thus undesirable, immunogenicity. It appears that peptides smaller than 10 amino acids are problematic for receptor recognition by class II MHC molecules ([0108] Immunology, Fifth edition, ed. Roitt et al., 1998, Mosby International Ltd., London, pp88-89).
  • Based on this study of Aβ, the useful B cell site derived from Aβ[0109] 1-42 should be in the size range of about 10 to about 28 residues.
    • EXAMPLE 5 Site-Directed Immunoreactivity Targeted by the Synthetic Peptide Immunogen Linked to Artificial Th Epitope
  • The non-immunogenic N-terminal fragment such as Aβ[0110] 1-14 of Aβ peptide was linked either through an εN-lysine spacer to an artificial Th peptide designated as MVF Th 1-16 (SEQ ID NO:51), or through a standard chemical coupling procedure to a conventional carrier protein KLH. The two immunogenic constructs were evaluated in guinea pigs for their relative “site-directed” immunogenicities to Aβ peptide and the resultant respective reactivity of the antibodies towards their respective carriers, the artificial Th epitope or the KLH carrier protein, according to the procedures described in Example 2. The short Aβ1-14 peptide alone as a control immunogen, and the two immunogenic constructs were formulated in a water-in-oil emulsion containing the adjuvant ISA51, a formulation that is suitable for human use. As shown in Table 8, the N-terminal Aβ1-14 fragment by itself is non-immunogenic as expected. The synthetic immunogen comprising Aβ1-14 fragment and artificial Th (SEQ ID NO: 73) was found to be highly immunogenic in eliciting site-directed antibodies to Aβ1-14. The antibodies were also found to be highly cross-reactive to soluble Aβ1-42 peptide as early as 4 weeks after the initial immunization (Log10 titers of 4.094 and 4.126 for 4 and 6 weeks post initial immunization respectively). When these AP-reactive high titer immune sera were tested by ELISA on the MVF Th1-16 peptide (SEQ ID NO 51) coated plate, they were found to be negative (Log10 titer of 0.038 and 0.064 for 4 and 6 weeks post initial immunization respectively) showing that irrelevant antibodies were not produced. The data obtained as shown in Table 8 clearly demonstrated the highly specific site-directed characteristic of the peptide immunogen of the present invention.
  • The immunogens with the carrier protein KLH was found to be highly -; immunoreactive with the conventional peptide-carrier protein conjugate (e.g. Log[0111] 10 .: titers of 4.903 and 5.018 for 4 and 6 weeks post initial immunization respectively). However, the antibodies elicited were only moderately crossreactive with the soluble Aβ1-42 peptide (e.g. with Log10 titers of 3.342 and 2.736 for 4 and 6 weeks post initial immunization respectively). This is approximately 1× to 100× less than SEQ ID NO:73. Unexpectedly, the peptide immunogens of the present invention were highly site-directed and focused. Only functionally important antibodies towards the anti-aggregation and disaggregation sites on the N-terminal fragment of the Aβ peptide were generated rather than towards irrelevant carrier sites.
    • EXAMPLE 6 Evaluation of Aβ Peptide Immunogen by Cross-Reactivities to Senile Plaques
  • Brains of AD patients with plaques and tangles and thioflavine S positive blood vessels (TSBV) containing amyloid plaques were used for evaluation of cross-reactivities to polymeric senile plaques of the immune sera raised in guinea pigs and baboons against Aβ peptide immunogens. Plaques and TSBV reactivities were detected by immunoperoxidase staining using Avidin-Biotinylated antibody Complex (ABC) method or by immunofluorescence staining using rhodamine conjugated Fab fragment of species specific anti-IgG. All guinea pig sera were tested at a dilution of 1:100 with end point titers determined for some of the samples. All baboon sera were tested at a dilution of 1:50. The evaluation of the immune and preimmune sera were kindly performed under code by Dr. Gaskin as described (Gaskin et al., [0112] J. Exp Med. 165:245, 1987).
  • In FIG. 1, serial cross sections of brains from 2 AD patients were initially examined at 1× magnification. Sections (a), (b) and (c) are from AD Brain 1 and (d), (e) and (e are from AD brain 2. Preimmune normal serum and immune sera from guinea pigs collected at 6 weeks post-initial immunization were tested by immunoperoxidase staining on cryostat sections from AD temporal cortex rich in plaques and neurofilament tangles (NFT). The immune sera used in the first study shown on slides FIGS. 1[0113] a and 1 d were obtained from animals immunized with Aβ1-28-εK-MvF Th1-16 (SEQ ID NO:74) prepared in ISA51 water-in-oil emulsion. The results show significant binding to both senile plaques and amyloid plaques on the thioflavine S positive blood vessels (TSBV). The cross-reactivities of the immune sera raised against the equivalent immunogen prepared in CFA/ICFA are shown in slides FIGS. 1b and 1 d. Unexpectedly, in contrast to the results obtained with the vaccine formulated with ISA51, preferential binding to the Aβ1-28 plaques on the blood vessels (TSBV) were observed for the sera raised against the CFA/ICFA vaccine. This means that the antibodies elicited by the vaccine formulated with ISA51 is distinguishable from the antibodies raised by the vaccine formulated in CFA/ICFA. Moreover, the antibodies generated by the vaccines formulated according to the present invention provided antibodies that have the desired higher cross reactivity to senile plaques in the brain tissue. Preimmune serum gave no staining in corresponding serial sections shown in slides FIGS. 1c and 1 f.
  • Further Immunoperoxidase staining of serial cross sections of AD brain 1 with preimmune and immune sera at 1:100 dilution are shown in FIGS. 2[0114] a to 2 e at 40×magnification. The sera obtained from animals immunized with Aβ1-28-εK-MVF Th 1-16 (Seq ID NO:74) prepared in ISA 51 water-in-oil emulsion strongly stained the plaques forming a pattern of cores as shown in slides FIGS. 2a and 2 d. Again, surprisingly, staining with immune sera prepared against the corresponding CFA/ICFA formulation gave a different staining pattern in that reactivities with plaques were predominantly on the blood vessels as shown in FIG. 2b rather than with the plaques in the brain tissue. Preimmune serum did not stain the sections as shown in FIG. 2c. The hyperimmune sera generated by immunization with Aβ1-42 peptide alone in CFA/ICFA, despite its strong reactivities with Aβ1-28 by ELISA, gave a surprisingly weak staining pattern in the section shown in FIG. 2e.
  • Similar immunostaining of AD brain tissue was performed with 11 pooled immune and preimmune sera obtained from guinea pigs immunized with the various vaccine formulations described in Examples 3, 4 and 5. These sera were also evaluated for their antibody reactivities with the functional-site by Aβ[0115] 1-14 ELISA, and with the soluble Aβ1-42 by Aβ1-42 ELISA (Table 9). In general, parallel trends were found with sera tested in all three assays. As shown in Table 9, the anti-peptide reactivities of the pre-immune serum and the sera raised against the short peptide Aβ1-14 alone formulated in ISA51 water-in-oil emulsion by ELISA were low and the cross-reactivities to plaques were negligible. Modest reactivities were found with sera from animals vaccinated with Aβ1-28 peptide alone formulated in Alum and in ISA51, and Aβ1-14 conjugated to KLH and formulated in ISA51. Whereas, significant site-directed reactivities to the functional Aβ1-14 site, to soluble Aβ, and to the plaques and TSBV in AD patient brain tissue sections were found with sera from animals immunized with synthetic Aβ/Th immunogens of the present invention. The results obtained from these studies, therefore, demonstrate excellent and useful immunogenicity of the peptide immunogens comprising the N-terminal fragment of Aβ1-42 having amino acids from 1-28 to about 1-10, linked to foreign Th epitopes. Moreover, the results showed that the presence of a foreign Th epitope improves the immunogenicity of the peptide immunogens of the present invention to a surprising extent. The peptide immunogens of the present invention in clinically acceptable vaccine formulations acceptable to use in humans generated antibodies having the desired cross-reactivity to senile plaques in the brain tissues of AD patients.
    • EXAMPLE 7 The Immunogenicity of Representative Aβ Peptide Vaccines in Baboons as Predictor of Immunotherapeutic Efficacy for AD
  • A representative synthetic immunogen, Aβ[0116] 1-28-ε-K-MvF Th1-16 (SEQ ID NO:74), formulated in ISA51 water-in-oil emulsion at dose levels of 25 ug/0.5 mL, 100 ug/0.5 mL and 400 ug/0.5 mL were given to three baboons Y299, X398, X1198 at 0, 3 and 6 weeks schedule from initial immunization. Pre-immune sera and sera at weeks 5 and 8 weeks post initial immunization (wpi) were collected. For comparison, a fourth baboon X798 was given 100 ug/0.5 mL doses of an equimolar mixture of free peptides Aβ1-28 and Aβ1-42 formulated in alum, the standard adjuvant approved for human use. Preimmune sera were used as the negative control.
  • Sera from all four immunized animals were collected and evaluated for their antibody reactivities with the functional site by A[0117] 1-14 ELISA, and for reactivities with soluble Aβ1-42 by Aβ1-42 ELISA (for sera collected at 0, 5 and 8 wpi). The cross-reactivities of the anti-sera (8 wpi only) with the senile plaques and the plaques in thioflavine S positive blood vessels were evaluated by immunostaining as described in Example 6. Instead of using anti-baboon Ig, the antibody detector used is an Fab fragment from anti-human IgG that recognizes all human isotypes and is cross-reactive with baboon IgG.
  • Parallel trends again were found with sera tested in all three assays. As shown in Table 10, pre-immune sera were negative. Modest ELISA reactivities were found with serum from animal X798 vaccinated with Aβ[0118] 1-28 and Aβ1-42 formulated in Alum. However, the reactivity of this serum was weak for the recognition of senile plaques. In contrast, significant site-directed reactivities to the functional-site of Aβ1-14, to soluble Aβ1-42, and to the plaques and TSBV in AD patient brain sections were found with sera collected at 8 weeks post initial immunization from animals immunized with the representative composition of the invention (SEQ ID NO:74) at both the 100 ug/0.5 mL and 400 ug/0.5 mL doses formulated with ISA51. The results obtained from this baboon study, therefore, demonstrated the usefulness of the immunogen of the present invention in a vaccine formulation appropriate for humans. The improvement in immunogenicity (10 to 100× increase in specific antibody titers to the functional-site of AP) is very significant in comparison to the peptide vaccine of the prior art with the immune responsiveness in baboons closely resembling that of humans.
  • Similarly, a mixture containing two to three synthetic immunogens of the present invention can be used for formulation into vaccines at from about 25 to 1000 ug per dose to elicit functional anti-Aβ[0119] 1-14 antibodies in genetically diverse human populations for the prevention and treatment of AD. Broad immunogenicity in humans is expected due to the presence of a promiscuous Th epitope in the peptide immunogen of the invention that provides for achieving broad MHC recognition.
    TABLE 1
    Pathogen-derived Promiscuous T Helper Cell Epitopes (Th)
    Description of Th Amino Acid Sequence SEQ ID NO
    HBs Tha FFLLTRILTIPQSLD 1
    PT1 Tha KKLRRLLYMIYMSGLAVRVHVSKEEQYYDY 2
    TT1 Tha KKQYIKANSKFIGITEL 3
    TT2 Tha KKFNNFTVSFWLRVPKVSASHL 4
    PTIA Th a YMSGLAVRVHVSKEE 5
    TT3 Tha YDPNYLRTDSDKDRFLQTMVKLFNRIK 6
    PT2 Tha GAYARCPNGTRALTVAELRGNAEL 7
    MVF1 Tha LSEIKGVIVHRLEGV 8
    MVF2 Tha GILESRGI KARITHVDTESY 9
    TT4 Tha WVRDIIDDFTNESSQKT 10
    TT5 Tha DVSTIVPYIGPALNHV 11
    CT Tha ALNIWDRFDVFCTLGATTGYLKGNS 12
    DT1 Tha DSETADNLEKTVAALSILPGHGC 13
    DT2 Tha EEIVAQSIALSSLMVAQAIPLVGELVDIGFAATNFVESC 14
    PF Tha DHEKKHAKMEKASSVFNVVNS 15
    SM Tha KWFKTNAPNGVDEKHRH 16
    TraT1 Tha GLQGKHADAVKAKG 17
    TraT2 Tha GLAAGLCGMAADAMVEDVN 18
    TraT3 Tha STETGNQHHYQTRVVSNANK 19
    HBc50-69 b SDFFPSVRDLLDTASALYRE 20
    CTP11 Thc TINKPKGYVGKE 21
  • [0120]
    TABLE 2
    Artificial Idealized Th and Combinatorial Library Idealized Artificial Th
    a. MVF Th and Th epitopes derived therefrom
    Th Identifier Amino Acid Sequence SEQ ID NO
    MVF Th1 LSEIKGVIVHRLEGV 22
    SSAL1 Th1 DLSDLKGLLLHKLDGL 23
    EI EIR III RIE I 24
    V V VVV V V 25
    F F FFF F F 26
    MVF Th1-1 ISEIKGVIVHKIEGI 27
    MT RT TRM TM 28
    L L V 29
    MVF Th1-2 ISEIKGVIVHKIEGI 30
    T RT TR T 31
    MVF Th1-3 MSEIKGVIVHKLEGM 32
    LT MRT TRM TV 33
    MVF Th1-4 ISEIKGVIVHKIEGI 34
    MVF Th1-5 ITEIRTVIVTRIETI 35
    MVF Th1-6 MSEMKGVIVHKMEGM 36
    MVF Th1-7 LTEIRTVIVTRLETV 37
    MVF Th1-8 ISISEIKGVIVHKIEGILF 38
    MT RT TRM TM 39
    L L V 40
    MVF Th1-9 ISISEIKGVIVHKIEGILF 41
    T RT TR T 42
    MVF Th1-10 ISLSEIKGVIVHKLEGMLF 43
    MT MRT TRM TV 44
    MVF Th1-11 ISLTEIRTVIVTRLETVLF 45
    I I I 46
    MVF Th1-12 ISISEIKGVIVHKIEGILF 47
    MVF Th1-13 ISITEIRTVIVTRIETILF 48
    MVF Th1-14 ISMSEMKGVIVHKMEGMLF 49
    MVF Th1-15 ISLTEIRTVIVTRLETVLF 50
    MVF Th1-16 ISITEIKGVIVHRIETILF 51
    b. HBsAg Th, Prototype and Derivatives
    HbsAg-Th1 FFLLTRILTIPQSLD 52
    HbsAg-Th1-1 KKKFFLLTRILTIPQSLD 53
    HbsAg-Th1-2 FFLLTRILTIPQSL 54
    SSAL2 Th2 KKKLFLLTKLLTLPQSLD 55
    RRRIKII RII I L IR 56
    VRVV VV V I V 57
    FFF FFFVF 58
    F 59
    HbsAg Th1-3 KKKIITITRIITIITTID 60
    HbsAg Th1-4 KKKIITITRIITIITTI 61
    HbsAg Th1-5 KKKMMTMTRMITMITTID 62
    HbsAg Th1-6 FITMDTKFLLASTHIL 63
    HbsAg Th1-7 KKKFITMDTKFLLASTHIL 64
  • [0121]
    TABLE 3
    Amino Acid Sequences of Aβ1-42 Peptides and its N-terminus Fragments
    SEQ ID NO Amino Acid Sequence
    SEQ ID NO:65 Aβ1-42 DAEFRHDSGYEVHHQKLVFFAEDVGSNKGAIIGLMVGGVVIA
    SEQ ID NO:66 Aβ1-28 DAEFRHDSGYEVHHQKLVFFAEDVGSNK
    SEQ ID NO:67 Aβ1-14 DAEFRHDSGYEVHH
    SEQ ID NO:68 Aβ1-12 DAEFRHDSGYEV
    SEQ ID NO:69 Aβ1-10 DAEFRHDSGY
  • [0122]
    TABLE 4
    SEQ ID
    Immunogen Amino Acid Sequence NO
    Aβ1-28-GG-HBV DAEFRMDSGYEVHHQKLVFFAEDVGSNK-GG-FFLLTRILTIPQSLD 70
    Th
    Aβ1-10-εK-IS-MVF DAEFRHDSGY-εK-ISITEIKGVIVHRIETILF 71
    Th1-16
    Aβ1-12-εK-IS-MVF DAEFRHDSGYEV-εK-ISITEIKGVIVHRIETILF 72
    Th1-16
    Aβ1-14-εK-IS-MVF DAEFRHDSGYEVHH-εK-ISITEIKGVIVHRIETILF 73
    Th1-16
    Aβ1-14-εK-IS-MVF DAEFRHDSGYEVHHQKLVFFAEDVGSNK-εK-ISITEIKGVIVHRIETILF 74
    Th1-16
    Aβ1-14-εK-MVF DAEFRHDSGYEVHH-εK-ISISEIKGVIVHKIEGILF 75
    Th1-9 T RT TR T 76
  • [0123]
    TABLE 5
    ELISA Titer (Log10)
    4 WPI 6 WPI
    Immunogen Adjuvant GP ID # Aβ1-14 Avg. Aβ1-42 Avg. Aβ1-14 Avg. Aβ1-42 Avg.
    Aβ1-28 Alum 1630 1.244 2.326 0.878 2.401 0.888 1.966 1.202 2.405
    (SEQ ID NO:66) 1631 3.408 3.924 3.044 3.608
    Aβ1-42 Alum 1634 0.773 1.124 0.680 1.461 1.062 1.784 1.203 1.807
    (SEQ ID NO:65) 1635 1.474 2.242 2.505 2.510
  • [0124]
    TABLE 6
    ELISA Titer (Log10)
    GP 4 WPI 6 WPI
    Immunogen Adjuvant ID # Aβ1-14 Avg. Aβ1-42 Avg. Aβ1-14 Avg. Aβ1-42 Avg.
    Aβ1-14 ISA51 1658 1.168 1.129 1.229 0.975 1.100 1.271 1.285 1.080
    (SEQ ID NO: 67) 1659 1.090 0.720 1.441 0.874
    Aβ1-28 ISA51 1632 2.341 2.291 3.656 3.382 2.276 2.715 3.359 3.455
    (SEQ ID NO: 66) 1633 2.241 3.107 3.153 3.550
    Aβ1-28-GG-HBVTh ISA51 1642 4.792 4.612 4.526 4.582 4.548 4.498 4.441 4.261
    (SEQ ID NO: 70) 1643 4.432 4.637 4.447 4.081
    Aβ1-42 ISA51 1636 2.724 1.864 3.603 2.402 2.286 1.997 3.250 2.873
    (SEQ ID NO: 65) 1637 1.004 1.201 1.707 2.495
  • [0125]
    TABLE 7
    ELISA Titer (Log10)
    GP 4 WPI 6 WPI
    Immunogen Adjuvant ID # Aβ1-14 Avg. Aβ1-42 Avg. Aβ1-14 Avg. Aβ1-42 Avg.
    Aβ1-10-εK-MVF CFA/IFA 1666 4.293 4.495 4.924 5.087 4.414 4.320 5.180 5.265
    Th1-16 1667 4.696 5.250 4.225 5.350
    (SEQ ID NO: 71)
    Aβ1-12-εK-MVF CFA/IFA 1664 4.577 4.495 5.100 4.891 5.320 4.545 6.000 5.278
    Th1-16 1665 4.322 4.682 3.700 4.555
    (SEQ ID NO: 72)
    Aβ1-14-εK-MVF CFA/IFA 1660 3.700 3.285 4.677 5.060 4.544 4.683 5.250 5.625
    Th1-16 1661 4.764 5.443 4.822 6.000
    (SEQ ID NO: 73)
    Aβ1-28-εK-MVF CFA/IFA 1584 3.355 3.201 4.610 4.328 2.743 3.592 4.487 4.901
    Th1-16 1585 3.707 4.688 3.731 5.155
    (SEQ ID NO: 74) 1586 2.545 3.685 4.304 5.061
  • [0126]
    TABLE 8
    ELISA Titer (Log10)
    4 WPI 6 WPI
    Th Th
    GP ID peptide peptide
    Immunogen Adjuvant # Aβ1-42 Avg or KLH Avg. Aβ1-42 Avg. or KLH Avg
    Aβ1-14 ISA 51 1658 1.229 0.975 NA NA 1.285 1.080 NA NA
    (SEQ ID NO: 67) 1659 0.720 NA 0.874 NA
    Aβ1-14-εK-MVF ISA 51 1662 4.388 4.094 0.006 0.038 4.559 4.261 0.065 0.064
    Th1-16 1663 3.800 0.070 3.693 0.063
    (SEQ ID NO: 73)
    KLH-(C) Aβ1-14 ISA 51 1670 3.181 3.342 4.672 4.903 2.625 2.736 4.876 5.018
    (SEQ ID NO: 67) 1671 3.502 5.133 2.846 5.160
  • [0127]
    TABLE 9
    Immunostaininga of
    serial frozen
    ELISA Titer (Log10) sections of AD's
    GP Aβ1-42 Aβ1-14 brain tissue
    Vaccine Formulation ID # Avg Avg Plaque TSBV
    Aβ1-28 in Alum 1630 0.878 2.401 1.244 2.326 +1 +4
    1631 3.924 3.408
    Aβ1-28 in ISA51 1632 3.686 3.397 2.341 2.291 +3 +5
    1633 3.107 2.241
    Aβ1-28-εK-MVF Th1-16 in CFM/IFA 1584 4.610 4.328 3.355 3.201 +4 +6
    (SEQ ID NO: 74) 1585 4.688 3.707
    1586 3.685 2.540
    Aβ1-28-εK-MVF Th1-16 in ISA51 1642 3.603 4.582 2.724 3.510 +4 +6
    (SEQ ID NO: 74) 1643 1.201 1.004
    Aβ1-14 in ISA51 1658 1.229 0.975 1.168 1.129 Neg Neg
    1659 0.720 1.090
    Aβ1-14-εK-MVF Th1-16 in CFA/IFA 1660 4.677 5.060 3.700 4.232 +4 +6
    (SEQ ID NO: 73) 1661 5.443 4.764
    Aβ1-14-εK-MVF Th1-16 in ISA51 1662 4.388 4.094 3.551 3.285 +4 +6
    (SEQ ID NO: 73) 1663 3.800 3.018
    Aβ1-12-εK-MVF Th1-16 in CFA/IFA 1664 5.100 4.891 4.577 4.450 +4 +6
    (SEQ ID NO: 72) 1665 4.682 4.322
    Aβ1-10-εK-MVF Th1-16 in CFA/IFA 1666 4.924 5.087 4.293 4.455 +4 +5
    (SEQ ID NO: 71) 1667 5.250 4.696
    KLH-(C) Aβ1-14 in ISA51 1670 3.181 3.342 3.280 3.102 +2 +4
    1635 3.502 2.924
    Negative Control <0.5 <0.5 Neg Neg
    (preimmune serum)
  • [0128]
    TABLE 10
    ELISA Titer (Log10) Immunostaining of
    Aβ1-42 Aβ1-14 frozen sections of
    Group 0 5 8 0 5 8 AD brain (8 wpi)
    # Vaccine Formulation Dose WPI WPI WPI WPI WPI WPI Plaques TSBV
    1 Aβ1-28-εKV-MVF Th1-16  25 μg 0.894 2.962 2.736 0.665 1.745 2.706 +2 +
    2 in ISA51 100 μg 0.610 2.987 3.640 0.794 2.816 4.800 +4 +6
    3 400 μg 0.696 2.696 4.050 0.539 4.250 3.799 +4 +6
    4 Aβ1-28+ Aβ1-42 in Alum 100 μg 0.897 1.963 2.485 0.798 0.727 2.850 + +
    5 Negative control — — — — — — — Neg Neg
  • [0129]
  • 1 76 1 15 PRT Hepatitis B virus 1 Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp 1 5 10 15 2 30 PRT Bordetella pertussis 2 Lys Lys Leu Arg Arg Leu Leu Tyr Met Ile Tyr Met Ser Gly Leu Ala 1 5 10 15 Val Arg Val His Val Ser Lys Glu Glu Gln Tyr Tyr Asp Tyr 20 25 30 3 17 PRT Clostridium tetani 3 Lys Lys Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile Gly Ile Thr Glu 1 5 10 15 Leu 4 22 PRT Clostridium tetani 4 Lys Lys Phe Asn Asn Phe Thr Val Ser Phe Trp Leu Arg Val Pro Lys 1 5 10 15 Val Ser Ala Ser His Leu 20 5 15 PRT Bordetella pertussis 5 Tyr Met Ser Gly Leu Ala Val Arg Val His Val Ser Lys Glu Glu 1 5 10 15 6 27 PRT Clostridium tetani 6 Tyr Asp Pro Asn Tyr Leu Arg Thr Asp Ser Asp Lys Asp Arg Phe Leu 1 5 10 15 Gln Thr Met Val Lys Leu Phe Asn Arg Ile Lys 20 25 7 24 PRT Pertusaria trachythallina 7 Gly Ala Tyr Ala Arg Cys Pro Asn Gly Thr Arg Ala Leu Thr Val Ala 1 5 10 15 Glu Leu Arg Gly Asn Ala Glu Leu 20 8 15 PRT Measles virus 8 Leu Ser Glu Ile Lys Gly Val Ile Val His Arg Leu Glu Gly Val 1 5 10 15 9 20 PRT Escherichia coli 9 Gly Ile Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile Thr His Val Asp 1 5 10 15 Thr Glu Ser Tyr 20 10 17 PRT Clostridium tetani 10 Trp Val Arg Asp Ile Ile Asp Asp Phe Thr Asn Glu Ser Ser Gln Lys 1 5 10 15 Thr 11 16 PRT Clostridium tetani 11 Asp Val Ser Thr Ile Val Pro Tyr Ile Gly Pro Ala Leu Asn His Val 1 5 10 15 12 25 PRT Chlamydia trachomatis 12 Ala Leu Asn Ile Trp Asp Arg Phe Asp Val Phe Cys Thr Leu Gly Ala 1 5 10 15 Thr Thr Gly Tyr Leu Lys Gly Asn Ser 20 25 13 23 PRT Diphtheria 13 Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Val Ala Ala Leu Ser 1 5 10 15 Ile Leu Pro Gly His Gly Cys 20 14 39 PRT Diphtheria 14 Glu Glu Ile Val Ala Gln Ser Ile Ala Leu Ser Ser Leu Met Val Ala 1 5 10 15 Gln Ala Ile Pro Leu Val Gly Glu Leu Val Asp Ile Gly Phe Ala Ala 20 25 30 Thr Asn Phe Val Glu Ser Cys 35 15 21 PRT Plasmodium falciparum 15 Asp His Glu Lys Lys His Ala Lys Met Glu Lys Ala Ser Ser Val Phe 1 5 10 15 Asn Val Val Asn Ser 20 16 17 PRT Schistosoma mansoni 16 Lys Trp Phe Lys Thr Asn Ala Pro Asn Gly Val Asp Glu Lys His Arg 1 5 10 15 His 17 14 PRT Escherichia coli 17 Gly Leu Gln Gly Lys His Ala Asp Ala Val Lys Ala Lys Gly 1 5 10 18 19 PRT Escherichia coli 18 Gly Leu Ala Ala Gly Leu Val Gly Met Ala Ala Asp Ala Met Val Glu 1 5 10 15 Asp Val Asn 19 20 PRT Escherichia coli 19 Ser Thr Glu Thr Gly Asn Gln His His Tyr Gln Thr Arg Val Val Ser 1 5 10 15 Asn Ala Asn Lys 20 20 20 PRT Hepatitis B virus 20 Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr Ala Ser Ala 1 5 10 15 Leu Tyr Arg Glu 20 21 12 PRT Chlamydia trachomatis 21 Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu 1 5 10 22 15 PRT Measles virus 22 Leu Ser Glu Ile Lys Gly Val Ile Val His Arg Leu Glu Gly Val 1 5 10 15 23 16 PRT Measles virus 23 Asp Leu Ser Asp Leu Lys Gly Leu Leu Leu His Lys Leu Asp Gly Leu 1 5 10 15 24 16 PRT Measles virus 24 Glu Ile Ser Glu Ile Arg Gly Ile Ile Ile His Arg Ile Glu Gly Ile 1 5 10 15 25 16 PRT Measles virus 25 Asp Val Ser Asp Val Lys Gly Val Val Val His Lys Val Asp Gly Val 1 5 10 15 26 16 PRT Measles virus 26 Asp Phe Ser Asp Phe Lys Gly Phe Phe Phe His Lys Phe Asp Gly Phe 1 5 10 15 27 15 PRT Measles virus 27 Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly Ile 1 5 10 15 28 15 PRT Measles virus 28 Met Thr Glu Ile Arg Thr Val Ile Val Thr Arg Met Glu Thr Met 1 5 10 15 29 15 PRT Measles virus 29 Leu Ser Glu Ile Lys Gly Val Ile Val His Lys Leu Glu Gly Val 1 5 10 15 30 15 PRT Measles virus 30 Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly Ile 1 5 10 15 31 15 PRT Measles virus 31 Ile Thr Glu Ile Arg Thr Val Ile Val Thr Arg Ile Glu Thr Ile 1 5 10 15 32 15 PRT Measles virus 32 Met Ser Glu Ile Lys Gly Val Ile Val His Lys Leu Glu Gly Met 1 5 10 15 33 15 PRT Measles virus 33 Leu Thr Glu Met Arg Thr Val Ile Val Thr Arg Met Glu Thr Val 1 5 10 15 34 15 PRT Measles virus 34 Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly Ile 1 5 10 15 35 15 PRT Measles virus 35 Ile Thr Glu Ile Arg Thr Val Ile Val Thr Arg Ile Glu Thr Ile 1 5 10 15 36 15 PRT Measles virus 36 Met Ser Glu Met Lys Gly Val Ile Val His Lys Met Glu Gly Met 1 5 10 15 37 15 PRT Measles virus 37 Leu Thr Glu Ile Arg Thr Val Ile Val Thr Arg Leu Glu Thr Val 1 5 10 15 38 19 PRT Measles virus 38 Ile Ser Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly 1 5 10 15 Ile Leu Phe 39 19 PRT Measles virus 39 Ile Ser Met Thr Glu Ile Arg Thr Val Ile Val Thr Arg Met Glu Thr 1 5 10 15 Met Leu Phe 40 19 PRT Measles virus 40 Ile Ser Leu Ser Glu Ile Lys Gly Val Ile Val His Lys Leu Glu Gly 1 5 10 15 Val Leu Phe 41 19 PRT Measles virus 41 Ile Ser Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly 1 5 10 15 Ile Leu Phe 42 19 PRT Measles virus 42 Ile Ser Ile Thr Glu Ile Arg Thr Val Ile Val Thr Arg Ile Glu Thr 1 5 10 15 Ile Leu Phe 43 19 PRT Measles virus 43 Ile Ser Leu Ser Glu Ile Lys Gly Val Ile Val His Lys Leu Glu Gly 1 5 10 15 Met Leu Phe 44 19 PRT Measles virus 44 Ile Ser Met Thr Glu Met Arg Thr Val Ile Val Thr Arg Met Glu Thr 1 5 10 15 Val Leu Phe 45 19 PRT Measles virus 45 Ile Ser Leu Thr Glu Ile Arg Thr Val Ile Val Thr Arg Leu Glu Thr 1 5 10 15 Val Leu Phe 46 19 PRT Measles virus 46 Ile Ser Ile Thr Glu Ile Arg Thr Val Ile Val Thr Arg Ile Glu Thr 1 5 10 15 Ile Leu Phe 47 19 PRT Measles virus 47 Ile Ser Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly 1 5 10 15 Ile Leu Phe 48 19 PRT Measles virus 48 Ile Ser Ile Thr Glu Ile Arg Thr Val Ile Val Thr Arg Ile Glu Thr 1 5 10 15 Ile Leu Phe 49 19 PRT Measles virus 49 Ile Ser Met Ser Glu Met Lys Gly Val Ile Val His Lys Met Glu Gly 1 5 10 15 Met Leu Phe 50 19 PRT Measles virus 50 Ile Ser Leu Thr Glu Ile Arg Thr Val Ile Val Thr Arg Leu Glu Thr 1 5 10 15 Val Leu Phe 51 19 PRT Measles virus 51 Ile Ser Ile Thr Glu Ile Lys Gly Val Ile Val His Arg Ile Glu Thr 1 5 10 15 Ile Leu Phe 52 15 PRT Hepatitis B virus 52 Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp 1 5 10 15 53 18 PRT Hepatitis B virus 53 Lys Lys Lys Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser 1 5 10 15 Leu Asp 54 14 PRT Hepatitis B virus 54 Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu 1 5 10 55 18 PRT Hepatitis B virus 55 Lys Lys Lys Leu Phe Leu Leu Thr Lys Leu Leu Thr Leu Pro Gln Ser 1 5 10 15 Leu Asp 56 18 PRT Hepatitis B virus 56 Arg Arg Arg Ile Lys Ile Ile Thr Arg Ile Ile Thr Ile Pro Leu Ser 1 5 10 15 Ile Arg 57 18 PRT Hepatitis B virus 57 Lys Lys Lys Val Arg Val Val Thr Lys Val Val Thr Val Pro Ile Ser 1 5 10 15 Val Asp 58 18 PRT Hepatitis B virus 58 Lys Lys Lys Phe Phe Phe Phe Thr Lys Phe Phe Thr Phe Pro Val Ser 1 5 10 15 Phe Asp 59 18 PRT Hepatitis B virus 59 Lys Lys Lys Leu Phe Leu Leu Thr Lys Leu Leu Thr Leu Pro Phe Ser 1 5 10 15 Leu Asp 60 18 PRT Hepatitis B virus 60 Lys Lys Lys Ile Ile Thr Ile Thr Arg Ile Ile Thr Ile Ile Thr Thr 1 5 10 15 Ile Asp 61 17 PRT Hepatitis B virus 61 Lys Lys Lys Ile Ile Thr Ile Thr Arg Ile Ile Thr Ile Ile Thr Thr 1 5 10 15 Ile 62 18 PRT Hepatitis B virus 62 Lys Lys Lys Met Met Thr Met Thr Arg Met Ile Thr Met Ile Thr Thr 1 5 10 15 Ile Asp 63 16 PRT Hepatitis B virus 63 Phe Ile Thr Met Asp Thr Lys Phe Leu Leu Ala Ser Thr His Ile Leu 1 5 10 15 64 19 PRT Hepatitis B virus 64 Lys Lys Lys Phe Ile Thr Met Asp Thr Lys Phe Leu Leu Ala Ser Thr 1 5 10 15 His Ile Leu 65 42 PRT Homo sapiens 65 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln Lys 1 5 10 15 Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Gly Ala Ile Ile 20 25 30 Gly Leu Met Val Gly Gly Val Val Ile Ala 35 40 66 28 PRT Homo sapiens 66 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln Lys 1 5 10 15 Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys 20 25 67 14 PRT Homo sapiens 67 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His 1 5 10 68 12 PRT Homo sapiens 68 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val 1 5 10 69 10 PRT Homo sapiens 69 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr 1 5 10 70 45 PRT Hepatitis B virus 70 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln Lys 1 5 10 15 Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Gly Gly Phe Phe 20 25 30 Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp 35 40 45 71 30 PRT Measles virus 71 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Lys Ile Ser Ile Thr Glu 1 5 10 15 Ile Lys Gly Val Ile Val His Arg Ile Glu Thr Ile Leu Phe 20 25 30 72 32 PRT Measles virus 72 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val Lys Ile Ser Ile 1 5 10 15 Thr Glu Ile Lys Gly Val Ile Val His Arg Ile Glu Thr Ile Leu Phe 20 25 30 73 34 PRT Measles virus 73 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Lys Ile 1 5 10 15 Ser Ile Thr Glu Ile Lys Gly Val Ile Val His Arg Ile Glu Thr Ile 20 25 30 Leu Phe 74 48 PRT Measles virus 74 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Gln Lys 1 5 10 15 Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Lys Ile Ser Ile 20 25 30 Thr Glu Ile Lys Gly Val Ile Val His Arg Ile Glu Thr Ile Leu Phe 35 40 45 75 34 PRT Measles virus 75 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Lys Ile 1 5 10 15 Ser Ile Ser Glu Ile Lys Gly Val Ile Val His Lys Ile Glu Gly Ile 20 25 30 Leu Phe 76 34 PRT Measles virus 76 Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His Lys Ile 1 5 10 15 Ser Ile Thr Glu Ile Arg Thr Val Ile Val Thr Arg Ile Glu Thr Ile 20 25 30 Leu Phe

Claims (80)

I claim:
1. A peptide immunogen of about 20 to 100 amino acids long comprising:
(i) a helper T cell (Th) epitope selected from the group consisting of SEQ ID Nos: 1 to 64;
(ii) an N-terminal fragment of Aβ1-42 peptide, SEQ ID NO:65, consisting of from 10 to 28 amino acid residues wherein each fragment comprises amino acid residue 1 of the Aβ1-42 peptide or an immunologically functional analog of the N-teminal fragment of Aβ1-42 peptide; and
(iii) optionally a spacer consisting of at least an amino acid to separate the immunogenic domains.
2. A peptide immunogen of claim 1, wherein the spacer is selected from the group consisting of an amino acid, Gly-Gly, ((α, ε-N)-Lys, and Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:73).
3. A peptide immunogen of claim 2, wherein the spacer is Gly-Gly.
4. A peptide immunogen of claim 2, wherein the spacer is ε-N-Lys.
5. A peptide immunogen of claim 1, wherein the N-terminal fragment of Aβ1-42 peptide is selected from the group consisting of SEQ ID NOs: 66-69 and an immunologically functional analog thereof.
6. A peptide immunogen of any one of claims 2, 3, or 4, wherein the N-terminal fragment of Aβ1-42 peptide is selected from the group consisting of SEQ ID NOs: 66-69 and an immunologically functional analog thereof.
7. A peptide immunogen of claim 1, wherein Th is selected from the group consisting of SEQ ID NOs: 1, 3, 4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54.
8. A peptide immunogen of any one of claims 2, 3, or 4, wherein Th is selected from the group consisting of SEQ ID NOs: 1, 3, 4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54.
9. A peptide immunogen selected from the group consisting of SEQ ID NOs: 70, 71, 72, 73, and 74.
10. A peptide immunogen of claim 9 consisting of SEQ ID NO: 73.
11. A peptide immunogen of claim 9 consisting of SEQ ID NO: 74.
12. The peptide immunogen represented by one of the following formulae:
(A)n-(N-terminal fragment of Aβ1-42 peptide)-(B)o-(Th)m-X; or (A)n-(Th)m-(B)o-(N-terminal fragment of Aβ1-42 peptide)-X;
wherein
each A is independently an amino acid;
each B is a linking group selected from the group consisting of an amino acid, gly-gly, (α, ε-N)-Lys, and Pro-Pro-Xaa-Pro-Xaa-Pro (SEQ ID NO:73);
Th comprise an amino acid sequence that constitutes a helper T cell epitope, selected from the group consisting of SEQ ID NOs: 1-64 and an immune enhancing analog thereof;
(N-terminal fragment of Aβ1-42 peptide) is 10 to about 28 amino acid residues and wherein each fragment comprises EFRH of the Aβ1-42 peptide and immunologically functional analog thereof;
X is an α-COOH or α-CONH2 of an amino acid;
n is from 0 to about 10;
m is from 1 to about 4; and
o is from 0 to about 10.
13. A peptide immunogen of claim 12, wherein the spacer is Gly-Gly.
14. A peptide immunogen of claim 12, wherein the spacer is ε-N-Lys.
15. A peptide immunogen of claim 12, wherein the N-terminal fragment of Aβ1-42 peptide is selected from the group consisting of SEQ ID NOs: 66-69 and an immunologically effective analog thereof.
16. A peptide immunogen of any one of claims 13, or 14, wherein the N-terminal fragment of Aβ1-42 peptide is selected from the group consisting of SEQ ID NOs: 66-69 and an immunologically functional analog thereof
17. A peptide immunogen of claim 12, wherein Th is selected from the group consisting of SEQ ID NOs: 1, 3,4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54.
18. A peptide immunogen of any one of claims 13, or 14 wherein Th is selected from the group consisting of SEQ ID NOs: 1, 3, 4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54.
19. A peptide immunogen of claim 15 wherein Th is selected from the group consisting of SEQ ID NOs: 1, 3, 4, 5, 6, 7, 8, 9, 20, 38-40, 47-51 and 52-54.
20. A peptide immunogen of claim 16 wherein Th is selected from the group consisting of SEQ ID NOs: 1, 3,4, 5, 6, 7, 8, 9, 20, 38-40,47-51 and 52-54.
21. A composition comprising a pepetide immunogen of claim 1 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
22. A composition comprising a pepetide immunogen of claim 2 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
23. A composition comprising a pepetide immunogen of claim 3 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
24. A composition comprising a pepetide immunogen of claim 4 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
25. A composition comprising a pepetide immunogen of claim 5 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
26. A composition comprising a pepetide immunogen of claim 6 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
27. A composition comprising a pepetide immunogen of claim 7 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
28. A composition comprising a pepetide immunogen of claim 8 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
29. A composition comprising a pepetide immunogen of claim 9 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
30. A composition comprising a pepetide immunogen of claim 10 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
31. A composition comprising a pepetide immunogen of claim 11 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
32. A composition comprising a pepetide immunogen of claim 12 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720
33. A composition comprising a pepetide immunogen of claim 13 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
34. A composition comprising a pepetide immunogen of claim 14 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
35. A composition comprising a pepetide immunogen of claim 15 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
36. A composition comprising a pepetide immunogen of claim 16 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
37. A composition comprising a pepetide immunogen of claim 17 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
38. A composition comprising a pepetide immunogen of claim 18 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
39. A composition comprising a pepetide immunogen of claim 19 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, , ISA35, ISA206 and ISA 720.
40. A composition comprising a pepetide immunogen of claim 20 and a pharmaceutically acceptable adjuvant and/or carrier selected from the group consisting of alum, liposyn, saponin, squalene, L121, emulsigen monophosphyryl lipid A (MPL), polysorbate 80, QS21, Montamide ISA51, ISA35, ISA206 and ISA 720.
41. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 21.
42. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 22.
43. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 23.
44. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 24.
45. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 25.
46. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 26.
47. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 27.
48. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 28.
49. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 29.
50. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 30.
51. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 31.
52. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 32.
53. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 33.
54. A method of preventing Alzheimer's disease by administrating to a mammal a composition of claim 34.
55. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 35.
56. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 36.
57. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 37.
58. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 38.
59. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 39.
60. A method of preventing or treating Alzheimer's disease by administrating to a mammal a composition of claim 40.
61. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 21.
62. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 22.
63. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 23.
64. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 24.
65. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 25.
66. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 26.
67. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 27.
68. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 28.
69. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 29.
70. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 30.
71. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 31.
72. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 32.
73. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 33.
74. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 34.
75. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 35.
76. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 36.
77. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 37.
78. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 38.
79. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 39.
80. A method of producing antibodies to Aβ1-42 peptide that is cross reactive to soluble Aβ peptides and brain tissue plaques formed therefrom by administering a composition of claim 40.
US09/865,294 2001-05-25 2001-05-25 Immunogenic peptide composition comprising measles virus Fprotein Thelper cell epitope (MUFThl-16) and N-terminus of β-amyloid peptide Expired - Lifetime US6906169B2 (en)

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US09/865,294 US6906169B2 (en) 2001-05-25 2001-05-25 Immunogenic peptide composition comprising measles virus Fprotein Thelper cell epitope (MUFThl-16) and N-terminus of β-amyloid peptide
TW091105087A TWI252233B (en) 2001-05-25 2002-03-18 Immunogenic peptide composition for the prevention and treatment of Alzheimer's disease
CA2665748A CA2665748C (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
AT09168110T ATE496937T1 (en) 2001-05-25 2002-04-02 IMMUNOGENIC PEPTIDE COMPOSITION FOR THE PREVENTION AND TREATMENT OF ALZHEIMER'S DISEASE
EP02731223A EP1497313B1 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
AU2002303211A AU2002303211B2 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
JP2002592863A JP4440544B2 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for prevention and treatment of Alzheimer's disease
CA002448171A CA2448171A1 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
BR0210010-0 BRPI0210010B8 (en) 2001-05-25 2002-04-02 peptide immunogen, as well as composition and application comprising them
AT02731223T ATE474000T1 (en) 2001-05-25 2002-04-02 IMMUNOGENIC PEPTIDE COMPOSITION FOR THE PREVENTION AND TREATMENT OF ALZHEIMER'S DISEASE
PCT/US2002/010293 WO2002096350A2 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
MXPA03010631A MXPA03010631A (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease.
DK02731223.0T DK1497313T3 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of Alzheimer's disease
CN2008101692748A CN101372511B (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
EP09168110A EP2123671B1 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of Alzheimer's Disease
ES09168110T ES2360465T3 (en) 2001-05-25 2002-04-02 COMPOSITION OF IMMUNOGENIC PEPTIDE FOR THE PREVENTION AND TREATMENT OF ALZHEIMER'S DISEASE.
CN028106210A CN1568329B (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of Alzheimer's disease
ES02731223T ES2348975T3 (en) 2001-05-25 2002-04-02 IMMUNOGEN PEPTIDE COMPOSITION FOR THE PREVENTION AND TREATMENT OF ALZEHIMER'S DISEASE.
DE60237044T DE60237044D1 (en) 2001-05-25 2002-04-02 IMMUNOGENIC PEPTIDE COMPOSITION FOR THE PREVENTION AND TREATMENT OF ALZHEIMER DISEASE
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DE60239093T DE60239093D1 (en) 2001-05-25 2002-04-02 Immunogenic peptide composition for the prevention and treatment of Alzheimer's disease
ZA2003/08767A ZA200308767B (en) 2001-05-25 2003-11-11 Immunogenic peptide composition for the prevention and treatment of alzheimers disease
US10/861,614 US7951909B2 (en) 2001-05-25 2004-06-04 Immunogenic peptide composition comprising a promiscuous helper T cell epitope and an N-terminal fragment of Aβ1-42 peptide
HK09107043.3A HK1127898A1 (en) 2001-05-25 2005-03-08 A peptide composition comprising immunogenicity used for preventing and treating alzheimer's disease
HK05102035.8A HK1068215A1 (en) 2001-05-25 2005-03-08 Immunogenic peptide composition for the prevention and treatment of alzheimer's disease
AU2008202270A AU2008202270B2 (en) 2001-05-25 2008-05-22 Immunogenic peptide composition for the prevention and treatment of Alzheimer's disease
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JP2009123670A JP5300592B2 (en) 2001-05-25 2009-05-22 Immunogenic peptide composition for prevention and treatment of Alzheimer's disease
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Cited By (40)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020187157A1 (en) * 2000-02-21 2002-12-12 Jensen Martin Roland Novel method for down-regulation of amyloid
US20030165478A1 (en) * 2002-02-14 2003-09-04 Sokoll Kenneth K. Stabilized synthetic immunogen delivery system
US20030165496A1 (en) * 2000-12-06 2003-09-04 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US20040082762A1 (en) * 2002-03-12 2004-04-29 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US20040087777A1 (en) * 2000-12-06 2004-05-06 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US6743427B1 (en) 1997-12-02 2004-06-01 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6750324B1 (en) 1997-12-02 2004-06-15 Neuralab Limited Humanized and chimeric N-terminal amyloid beta-antibodies
US6761888B1 (en) 2000-05-26 2004-07-13 Neuralab Limited Passive immunization treatment of Alzheimer's disease
US20040166119A1 (en) * 1997-12-02 2004-08-26 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040171816A1 (en) * 1997-12-02 2004-09-02 Schenk Dale B. Humanized antibodies that recognize beta amyloid peptide
US6787637B1 (en) 1999-05-28 2004-09-07 Neuralab Limited N-Terminal amyloid-β antibodies
US20040219146A1 (en) * 1997-12-02 2004-11-04 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050059591A1 (en) * 1998-04-07 2005-03-17 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6913745B1 (en) 1997-12-02 2005-07-05 Neuralab Limited Passive immunization of Alzheimer's disease
US20060165682A1 (en) * 2004-12-15 2006-07-27 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US20060198851A1 (en) * 2004-12-15 2006-09-07 Guriq Basi Humanized Abeta antibodies for use in improving cognition
US20060257396A1 (en) * 2004-12-15 2006-11-16 Jacobsen Jack S Abeta antibodies for use in improving cognition
US20070041945A1 (en) * 2000-02-21 2007-02-22 Pharnexa A/S Novel method for down-regulation of amyloid
US20070154480A1 (en) * 1998-04-07 2007-07-05 Schenk Dale B Humanized antibodies that recognize beta amyloid peptide
US20080050367A1 (en) * 1998-04-07 2008-02-28 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
WO2008042024A2 (en) 2006-06-01 2008-04-10 Elan Pharmaceuticals, Inc. Neuroactive fragments of app
US20080267986A1 (en) * 2005-12-12 2008-10-30 Andrea Pfeifer Therapeutic vaccine
US20080292625A1 (en) * 2007-04-18 2008-11-27 Sally Schroeter Prevention and treatment of cerebral amyloid angiopathy
US20090142270A1 (en) * 2007-04-18 2009-06-04 Elan Pharma International Limited Prevention and treatment of cerebral amyloid angiopathy
US20100047262A1 (en) * 2001-08-20 2010-02-25 Peter Birk Rasmussen Novel method for down-regulation of amyloid
US7700751B2 (en) 2000-12-06 2010-04-20 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize β-amyloid peptide
US20100266505A1 (en) * 2007-10-17 2010-10-21 Wyeth Llc Immunotherapy regimes dependent on apoe status
US20100322932A1 (en) * 1998-05-21 2010-12-23 Alan Solomon Methods for amyloid removal using anti-amyloid antibodies
US7871615B2 (en) 2003-05-30 2011-01-18 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize beta amyloid peptide
US20110229413A1 (en) * 2006-04-18 2011-09-22 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
US20110256172A1 (en) * 2008-10-14 2011-10-20 The Regents Of The University Of Michigan Epitope-targeted anthrax vaccine
US20120321694A1 (en) * 2010-10-27 2012-12-20 Daniel Larocque Compositions and uses
US8512709B2 (en) 2008-10-16 2013-08-20 The Chemo-Sero-Therapeutic Research Institue Modified amyloid β peptide
US8613920B2 (en) 2007-07-27 2013-12-24 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
US9051364B2 (en) 2008-11-19 2015-06-09 Forschungszentrum Juelich Gmbh Composition for producing anti-amyloid beta peptide antibodies with D-peptides
US9067981B1 (en) 2008-10-30 2015-06-30 Janssen Sciences Ireland Uc Hybrid amyloid-beta antibodies
US20160068581A1 (en) * 2013-03-15 2016-03-10 United Biomedical, Inc. Peptide vaccine for prevention and immunotherapy of dementia of the alzheimer's type
EP3329932A1 (en) 2009-06-10 2018-06-06 New York University Immunological targeting of pathological tau proteins
WO2023034914A3 (en) * 2021-09-01 2023-04-13 Vaxxinity, Inc. Methods for the prevention and treatment of synucleinopathies
WO2024158596A1 (en) * 2023-01-26 2024-08-02 Vaxxinity, Inc. Tau and amyloid beta peptide immunogen compositions and related methods

Families Citing this family (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6906169B2 (en) * 2001-05-25 2005-06-14 United Biomedical, Inc. Immunogenic peptide composition comprising measles virus Fprotein Thelper cell epitope (MUFThl-16) and N-terminus of β-amyloid peptide
EP1572894B1 (en) * 2001-11-21 2016-04-13 New York University Synthetic immunogenic but non-deposit-forming polypeptides and peptides homologous to amyloid beta, prion protein, amylin, alpha synuclein, or polyglutamine repeats for induction of an immune response thereto
US20040001848A1 (en) * 2002-03-01 2004-01-01 Szu-Yi Chou Method of producing disease-specific antigens
WO2003089460A1 (en) * 2002-04-19 2003-10-30 The Governing Council Of The University Of Toronto Immunological methods and compositions for the treatment of alzheimer's disease
EP1911765A3 (en) * 2002-07-24 2008-04-23 Innogenetics N.V. Prevention, treatment and diagnosis of diseases associated with Beta-Amyloid formation and/or aggregation
DE10303974A1 (en) 2003-01-31 2004-08-05 Abbott Gmbh & Co. Kg Amyloid β (1-42) oligomers, process for their preparation and their use
JP4888876B2 (en) 2003-06-13 2012-02-29 田平 武 Recombinant adeno-associated virus vector for the treatment of Alzheimer's disease
EP1515158B1 (en) * 2003-09-09 2013-07-17 Esaote S.p.A. Ultrasound imaging method combined with the presence of contrast media in the body under examination
PL2336147T3 (en) 2003-12-17 2015-01-30 Janssen Alzheimer Immunotherap A beta immunogenic peptide carrier conjugates and methods of producing same
JP2007522119A (en) * 2004-01-28 2007-08-09 キュリックス エーピーエス Conjugates of amyloid protein as vaccines for amyloid-related diseases
CU23297A1 (en) * 2004-11-16 2008-07-24 Ct De Inmunologa A Molecular IMMUNOTHERAPY / 00UTICAL FORMULATIONS FOR THE INDUCTION OF BLOCKING AUTHORTIC BODIES OF THE INTERLEUCINE-2 UNION TO ITS RECEIVER. ITS USE IN THE TREATMENT OF CÃ NCER
GB0427267D0 (en) * 2004-12-13 2005-01-12 Maria Teresa De Magistris Peptide adjuvants
JP5486808B2 (en) 2005-11-30 2014-05-07 アッヴィ・インコーポレイテッド Monoclonal antibody against amyloid beta protein and use thereof
EP2289909B1 (en) 2005-11-30 2014-10-29 AbbVie Inc. Screening method, process for purifying of non-diffusible a-beta oligomers, selective antibodies against said non-diffusible a-beta oligomers and a process for manufacturing of said antibodies
GB0613977D0 (en) 2006-02-07 2006-08-23 Peptcell Ltd Peptide sequences and compositions
CN101421299A (en) 2006-02-22 2009-04-29 株式会社林原生物化学研究所 Peptide vaccine for inducing production of anti-amyloid- -peptide antibody
CN101058608B (en) * 2006-04-21 2011-02-23 杜如昱 Human anti-Abeta(1-32) amyloid antibody, purifying method and use thereof
US8455626B2 (en) 2006-11-30 2013-06-04 Abbott Laboratories Aβ conformer selective anti-aβ globulomer monoclonal antibodies
US8895004B2 (en) 2007-02-27 2014-11-25 AbbVie Deutschland GmbH & Co. KG Method for the treatment of amyloidoses
JP5349297B2 (en) * 2007-04-20 2013-11-20 一般財団法人化学及血清療法研究所 Method for enhancing peptide immune response
EP2106802A1 (en) 2008-04-02 2009-10-07 SIGMA-TAU Industrie Farmaceutiche Riunite S.p.A. Modified peptides as synthetic vaccines in amyloid-associated disease
CN102256620B (en) 2008-08-07 2016-11-16 莫西亚药物公司 For treating the immunotherapeutic composition of Alzheimer
US20110221240A1 (en) * 2010-03-11 2011-09-15 Ziming Shen Compact children's table and stool set
JP2013523182A (en) 2010-04-15 2013-06-17 アボット・ラボラトリーズ Amyloid beta-binding protein
EP2590673A4 (en) 2010-07-08 2016-03-30 United Biomedical Inc Designer peptide-based pcv2 vaccine
CN105348387B (en) 2010-08-14 2020-08-25 Abbvie 公司 Amyloid beta binding proteins
GB201113570D0 (en) 2011-08-05 2011-09-21 Glaxosmithkline Biolog Sa Vaccine
CN102775478B (en) * 2012-08-10 2013-06-19 申联生物医药（上海）有限公司 Foot and mouth disease virus antigen peptide and vaccine
JP2015527369A (en) 2012-08-21 2015-09-17 インスティテュート フォー モレキュラー メディシン， インコーポレイテッド Compositions and methods relating to diseases associated with deposition of amyloid, tau and α-synuclein
US10195257B2 (en) 2013-07-28 2019-02-05 Qantu Therapeutics, Inc. Vaccine formulations comprising quillaja desacylsaponins and beta amyloid peptides or tau protein to induce a Th2 immune response
TWI609026B (en) * 2014-03-14 2017-12-21 美國聯合生物醫學公司 Peptide vaccine for prevention and immunotherapy of dementia of the alzheimer's type
EP3638298A4 (en) * 2017-06-16 2021-05-05 United Neuroscience Peptide immunogens from the c-terminal end of alpha-synuclein protein and formulations thereof for treatment of synucleinopathies
CN111405908B (en) 2017-10-27 2024-10-25 联脑科学公司 TAU peptide immunogen constructs
BR112020013352A2 (en) * 2017-12-31 2020-12-01 United Biomedical, Inc. peptide immunogens and formulations thereof directed to membrane-bound ige for the treatment of ige-mediated allergic diseases
KR20210010873A (en) * 2018-05-04 2021-01-28 유비아이 아이피 홀딩스 Artificial indiscriminate T helper cell epitope facilitating targeted antibody production with a limited T cell inflammatory response
WO2020072428A1 (en) * 2018-10-01 2020-04-09 United Neuroscience Peptide immunogen constructs directed against dipeptide repeat proteins from c9orf72
CN110684122B (en) * 2019-10-29 2021-03-02 中国人民解放军军事科学院军事医学研究院 Recombinant Tau epitope chimeric polymer antigen, preparation method and application thereof
CN113603773B (en) * 2021-08-17 2023-05-09 中国医科大学附属第一医院 Monoclonal antibody 7B8 targeting amyloid protein, hybridoma cell strain secreting antibody and application
CN116063447B (en) * 2022-09-13 2023-11-03 北京湃德智健科技有限公司 Antigen polypeptide for detecting ADAP autoantibody and application thereof

Family Cites Families (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1991016819A1 (en) 1990-04-27 1991-11-14 Molecular Rx., Inc. Method and composition for treatment of central nervous systems disease states associated with abnormal amyloid beta protein
US5753624A (en) * 1990-04-27 1998-05-19 Milkhaus Laboratory, Inc. Materials and methods for treatment of plaquing disease
US5955317A (en) * 1993-01-25 1999-09-21 Takeda Chemical Industries, Ltd. Antibodies to β-amyloids or their derivatives and use thereof
ATE239797T1 (en) * 1993-01-25 2003-05-15 Takeda Chemical Industries Ltd ANTIBODIES TO BETA-AMYLOID OR DERIVATIVES THEREOF AND THEIR USE
WO1994025060A1 (en) * 1993-04-27 1994-11-10 Ladd Anna E Immunogenic lhrh peptide constructs and synthetic universal immune stimulators for vaccines
US5759551A (en) 1993-04-27 1998-06-02 United Biomedical, Inc. Immunogenic LHRH peptide constructs and synthetic universal immune stimulators for vaccines
AU7043894A (en) * 1993-05-28 1994-12-20 Miriam Hospital, The Composition and method for (in vivo) imaging of amyloid deposits
CA2175579A1 (en) 1993-10-26 1995-05-04 Chang Yi Wang Structured synthetic antigen libraries as diagnostics, vaccines and therapeutics
US7964192B1 (en) * 1997-12-02 2011-06-21 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidgenic disease
TWI239847B (en) 1997-12-02 2005-09-21 Elan Pharm Inc N-terminal fragment of Abeta peptide and an adjuvant for preventing and treating amyloidogenic disease
US6787523B1 (en) * 1997-12-02 2004-09-07 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6750324B1 (en) * 1997-12-02 2004-06-15 Neuralab Limited Humanized and chimeric N-terminal amyloid beta-antibodies
TWI229679B (en) 1998-06-20 2005-03-21 United Biomedical Inc Artificial T helper cell epitopes as immune stimulators for synthetic peptide immunogens
US6025468A (en) 1998-06-20 2000-02-15 United Biomedical, Inc. Artificial T helper cell epitopes as immune stimulators for synthetic peptide immunogens including immunogenic LHRH peptides
UA81216C2 (en) * 1999-06-01 2007-12-25 Prevention and treatment of amyloid disease
JP2003509020A (en) * 1999-09-03 2003-03-11 ラモット・ユニバーシティ・オーソリティ・フォー・アプライド・リサーチ・アンド・インダストリアル・ディベロップメント・リミテッド Pharmaceuticals, compositions, and uses thereof useful for diagnosis, treatment, and prevention of plaque forming diseases
ES2275570T3 (en) 1999-12-08 2007-06-16 Intellect Neurosciences, Inc. BETA CHEMERIC AMYLOID PEPTIDES.
US20030165481A1 (en) 2000-02-24 2003-09-04 Hersh Louis B. Amyloid peptide inactivating enzyme to treat Alzheimer's disease
US6906169B2 (en) * 2001-05-25 2005-06-14 United Biomedical, Inc. Immunogenic peptide composition comprising measles virus Fprotein Thelper cell epitope (MUFThl-16) and N-terminus of β-amyloid peptide

Cited By (112)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6936246B1 (en) 1997-12-02 2005-08-30 Neuralab Limited Passive immunization of ASCR for prion disorders
US20050048049A1 (en) * 1997-12-02 2005-03-03 Neuralab Limited Prevention and treatment of amyloidogenic disease
US7964192B1 (en) 1997-12-02 2011-06-21 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidgenic disease
US7893214B2 (en) 1997-12-02 2011-02-22 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize beta amyloid peptide
US8034339B2 (en) 1997-12-02 2011-10-11 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidogenic disease
US20050191292A1 (en) * 1997-12-02 2005-09-01 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6743427B1 (en) 1997-12-02 2004-06-01 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6750324B1 (en) 1997-12-02 2004-06-15 Neuralab Limited Humanized and chimeric N-terminal amyloid beta-antibodies
US20090069544A1 (en) * 1997-12-02 2009-03-12 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US20040166119A1 (en) * 1997-12-02 2004-08-26 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040171816A1 (en) * 1997-12-02 2004-09-02 Schenk Dale B. Humanized antibodies that recognize beta amyloid peptide
US20040171815A1 (en) * 1997-12-02 2004-09-02 Schenk Dale B. Humanized antibodies that recognize beta amyloid peptide
US6787139B1 (en) 1997-12-02 2004-09-07 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6787138B1 (en) 1997-12-02 2004-09-07 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20080281082A1 (en) * 1997-12-02 2008-11-13 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US6787140B1 (en) 1997-12-02 2004-09-07 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6787144B1 (en) 1997-12-02 2004-09-07 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040175394A1 (en) * 1997-12-02 2004-09-09 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6808712B2 (en) 1997-12-02 2004-10-26 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040219146A1 (en) * 1997-12-02 2004-11-04 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6818218B2 (en) * 1997-12-02 2004-11-16 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040228865A1 (en) * 1997-12-02 2004-11-18 Neuralab Limited Prevention and treatment of amyloidogenic disease
US8535673B2 (en) 1997-12-02 2013-09-17 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidogenic disease
US20080096818A1 (en) * 1997-12-02 2008-04-24 Elan Pharma International Limited Prevention and treatment of amyloidogenic disease
US8642044B2 (en) 1997-12-02 2014-02-04 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidogenic disease
US8034348B2 (en) 1997-12-02 2011-10-11 Janssen Alzheimer Immunotherapy Prevention and treatment of amyloidogenic disease
US20050013815A1 (en) * 1997-12-02 2005-01-20 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050019330A1 (en) * 1997-12-02 2005-01-27 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050019343A1 (en) * 1997-12-02 2005-01-27 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050031629A1 (en) * 1997-12-02 2005-02-10 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050191314A1 (en) * 1997-12-02 2005-09-01 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6866849B2 (en) 1997-12-02 2005-03-15 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6866850B2 (en) 1997-12-02 2005-03-15 Neuralab Limited Prevention and treatment of amyloidogenic disease
US7014855B2 (en) 1997-12-02 2006-03-21 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20060034858A1 (en) * 1997-12-02 2006-02-16 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6905686B1 (en) 1997-12-02 2005-06-14 Neuralab Limited Active immunization for treatment of alzheimer's disease
US20050142132A1 (en) * 1997-12-02 2005-06-30 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6913745B1 (en) 1997-12-02 2005-07-05 Neuralab Limited Passive immunization of Alzheimer's disease
US20060029611A1 (en) * 1997-12-02 2006-02-09 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050163788A1 (en) * 1997-12-02 2005-07-28 Neuralab Limited Prevention and treatment of amyloidogenic disease
US9051363B2 (en) 1997-12-02 2015-06-09 Janssen Sciences Ireland Uc Humanized antibodies that recognize beta amyloid peptide
US6982084B2 (en) 1997-12-02 2006-01-03 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6972127B2 (en) 1997-12-02 2005-12-06 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050196399A1 (en) * 1997-12-02 2005-09-08 Schenk Dale B. Prevention and treatment of amyloidogenic disease
US6946135B2 (en) 1997-12-02 2005-09-20 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6962707B2 (en) 1997-12-02 2005-11-08 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050249725A1 (en) * 1997-12-02 2005-11-10 Schenk Dale B Humanized antibodies that recognize beta amyloid peptide
US20050249727A1 (en) * 1997-12-02 2005-11-10 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050255122A1 (en) * 1997-12-02 2005-11-17 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050059591A1 (en) * 1998-04-07 2005-03-17 Neuralab Limited Prevention and treatment of amyloidogenic disease
US7790856B2 (en) 1998-04-07 2010-09-07 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize beta amyloid peptide
US20070154480A1 (en) * 1998-04-07 2007-07-05 Schenk Dale B Humanized antibodies that recognize beta amyloid peptide
US20080050367A1 (en) * 1998-04-07 2008-02-28 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US20100322932A1 (en) * 1998-05-21 2010-12-23 Alan Solomon Methods for amyloid removal using anti-amyloid antibodies
US8105594B2 (en) 1998-05-21 2012-01-31 Alan Solomon Methods for amyloid removal using anti-amyloid antibodies
US20050009150A1 (en) * 1998-11-30 2005-01-13 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US6787637B1 (en) 1999-05-28 2004-09-07 Neuralab Limited N-Terminal amyloid-β antibodies
US20020187157A1 (en) * 2000-02-21 2002-12-12 Jensen Martin Roland Novel method for down-regulation of amyloid
US20090092579A1 (en) * 2000-02-21 2009-04-09 Pharmexa A/S Novel method for down-regulation of amyloid
US7135181B2 (en) * 2000-02-21 2006-11-14 Pharmexa A/S Method for down-regulation of amyloid
US20070041945A1 (en) * 2000-02-21 2007-02-22 Pharnexa A/S Novel method for down-regulation of amyloid
US20090311281A1 (en) * 2000-02-21 2009-12-17 H. Lundbeck A/S Novel method for down-regulation of amyloid
US20050158304A1 (en) * 2000-05-26 2005-07-21 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040265301A1 (en) * 2000-05-26 2004-12-30 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20040247590A1 (en) * 2000-05-26 2004-12-09 Neuralab Limited Prevention and treatment of amyloidogenic disease
US6761888B1 (en) 2000-05-26 2004-07-13 Neuralab Limited Passive immunization treatment of Alzheimer's disease
US20040247591A1 (en) * 2000-05-26 2004-12-09 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20060121038A9 (en) * 2000-05-26 2006-06-08 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20050123544A1 (en) * 2000-05-26 2005-06-09 Neuralab Limited Prevention and treatment of amyloidogenic disease
US20030165496A1 (en) * 2000-12-06 2003-09-04 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US20040087777A1 (en) * 2000-12-06 2004-05-06 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US20060280743A1 (en) * 2000-12-06 2006-12-14 Neuralab Limited Humanized antibodies that recognize beta amyloid peptide
US7700751B2 (en) 2000-12-06 2010-04-20 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize β-amyloid peptide
US20100047262A1 (en) * 2001-08-20 2010-02-25 Peter Birk Rasmussen Novel method for down-regulation of amyloid
US8871212B2 (en) 2001-08-20 2014-10-28 H. Lundbeck A/S Amyloid-beta polypeptide vaccine
US8088388B2 (en) * 2002-02-14 2012-01-03 United Biomedical, Inc. Stabilized synthetic immunogen delivery system
US8084015B2 (en) 2002-02-14 2011-12-27 United Biomedical, Inc. Stabilized synthetic immunogen delivery system
US20030165478A1 (en) * 2002-02-14 2003-09-04 Sokoll Kenneth K. Stabilized synthetic immunogen delivery system
US20040009897A1 (en) * 2002-02-14 2004-01-15 Sokoll Kenneth K. Stabilized synthetic immunogen delivery system
US20040082762A1 (en) * 2002-03-12 2004-04-29 Elan Pharmaceuticals, Inc. Humanized antibodies that recognize beta amyloid peptide
US8128928B2 (en) 2002-03-12 2012-03-06 Wyeth Llc Humanized antibodies that recognize beta amyloid peptide
US20110142823A1 (en) * 2002-03-12 2011-06-16 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize beta amyloid peptide
US7871615B2 (en) 2003-05-30 2011-01-18 Janssen Alzheimer Immunotherapy Humanized antibodies that recognize beta amyloid peptide
US20060165682A1 (en) * 2004-12-15 2006-07-27 Guriq Basi Humanized antibodies that recognize beta amyloid peptide
US20060198851A1 (en) * 2004-12-15 2006-09-07 Guriq Basi Humanized Abeta antibodies for use in improving cognition
US8916165B2 (en) 2004-12-15 2014-12-23 Janssen Alzheimer Immunotherapy Humanized Aβ antibodies for use in improving cognition
US20060257396A1 (en) * 2004-12-15 2006-11-16 Jacobsen Jack S Abeta antibodies for use in improving cognition
US20080267986A1 (en) * 2005-12-12 2008-10-30 Andrea Pfeifer Therapeutic vaccine
US7807175B2 (en) * 2005-12-12 2010-10-05 Ac Immune Sa Therapeutic vaccine
US20090238840A1 (en) * 2005-12-12 2009-09-24 Ac Immune, S.A. Therapeutic vaccine
US8603487B2 (en) 2005-12-12 2013-12-10 Ac Immune S.A. Palmitoylated AB 1-15 in a liposome as a treatment for amyloid associated diseases
US8784810B2 (en) 2006-04-18 2014-07-22 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
US20110229413A1 (en) * 2006-04-18 2011-09-22 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
WO2008042024A2 (en) 2006-06-01 2008-04-10 Elan Pharmaceuticals, Inc. Neuroactive fragments of app
US20080292625A1 (en) * 2007-04-18 2008-11-27 Sally Schroeter Prevention and treatment of cerebral amyloid angiopathy
US20090142270A1 (en) * 2007-04-18 2009-06-04 Elan Pharma International Limited Prevention and treatment of cerebral amyloid angiopathy
US8003097B2 (en) 2007-04-18 2011-08-23 Janssen Alzheimer Immunotherapy Treatment of cerebral amyloid angiopathy
US8613920B2 (en) 2007-07-27 2013-12-24 Janssen Alzheimer Immunotherapy Treatment of amyloidogenic diseases
US9644025B2 (en) 2007-10-17 2017-05-09 Wyeth Llc Immunotherapy regimes dependent on ApoE status
US20100266505A1 (en) * 2007-10-17 2010-10-21 Wyeth Llc Immunotherapy regimes dependent on apoe status
EP2952524A1 (en) 2007-10-17 2015-12-09 Janssen Sciences Ireland UC Immunotherapy regimes dependent on apoe status
US20110256172A1 (en) * 2008-10-14 2011-10-20 The Regents Of The University Of Michigan Epitope-targeted anthrax vaccine
US8512709B2 (en) 2008-10-16 2013-08-20 The Chemo-Sero-Therapeutic Research Institue Modified amyloid β peptide
US9067981B1 (en) 2008-10-30 2015-06-30 Janssen Sciences Ireland Uc Hybrid amyloid-beta antibodies
US9051364B2 (en) 2008-11-19 2015-06-09 Forschungszentrum Juelich Gmbh Composition for producing anti-amyloid beta peptide antibodies with D-peptides
EP3329932A1 (en) 2009-06-10 2018-06-06 New York University Immunological targeting of pathological tau proteins
EP4218794A2 (en) 2009-06-10 2023-08-02 New York University Immunological targeting of pathological tau proteins
US20120321694A1 (en) * 2010-10-27 2012-12-20 Daniel Larocque Compositions and uses
US20160068581A1 (en) * 2013-03-15 2016-03-10 United Biomedical, Inc. Peptide vaccine for prevention and immunotherapy of dementia of the alzheimer's type
US20190194280A1 (en) * 2013-03-15 2019-06-27 United Biomedical, Inc. Peptide vaccine for prevention and immunotherapy of dementia of the alzheimer's type
WO2023034914A3 (en) * 2021-09-01 2023-04-13 Vaxxinity, Inc. Methods for the prevention and treatment of synucleinopathies
WO2024158596A1 (en) * 2023-01-26 2024-08-02 Vaxxinity, Inc. Tau and amyloid beta peptide immunogen compositions and related methods

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